Compounds and compositions as protein kinase inhibitors

ABSTRACT

The invention provides a novel class of compounds, pharmaceutical compositions comprising such compounds and methods of using such compounds to treat or prevent diseases or disorders associated with abnormal or deregulated kinase activity, particularly diseases or disorders that involve abnormal activation of B-Raf.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalPatent Applications 61/238,073, filed 28 Aug. 2009 and 61/313,039, filed11 Mar. 2010. The full disclosures of these applications areincorporated herein by reference in their entirety and for all purposes.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention provides a novel class of compounds, pharmaceuticalcompositions comprising such compounds and methods of using suchcompounds to treat or prevent diseases or disorders associated withabnormal or deregulated kinase activity, particularly diseases ordisorders that involve abnormal activation of B-Raf.

2. Background

The protein kinases represent a large family of proteins, which play acentral role in the regulation of a wide variety of cellular processesand maintaining control over cellular function. A partial, non-limiting,list of these kinases include: receptor tyrosine kinases such asplatelet-derived growth factor receptor kinase (PDGF-R), the nervegrowth factor receptor, trkB, Met, and the fibroblast growth factorreceptor, FGFR3; non-receptor tyrosine kinases such as Abl and thefusion kinase BCR-Abl, Lck, Csk, Fes, Bmx and c-src; andserine/threonine kinases such as B-Raf, sgk, MAP kinases (e.g., MKK4,MKK6, etc.) and SAPK2α, SAPK2β and SAPK3. Aberrant kinase activity hasbeen observed in many disease states including benign and malignantproliferative disorders as well as diseases resulting from inappropriateactivation of the immune and nervous systems.

The novel compounds of this invention inhibit the activity of B-Raf ormutant forms thereof (for example V600E) and are, therefore, expected tobe useful in the treatment of B-Raf-associated diseases.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides compounds of Formula I:

in which:

Y is selected from N and CR₆;

R₁ is selected from hydrogen, —X₁R_(8a), —X₁OX₂R_(8a),—X₁C(O)NR_(8a)R_(8b), —X₁NR_(8a)X₂R_(8b), —X₁NR_(8a)C(O)X₂OR_(8b),—X₁NR_(8a)C(O)X₂NR_(8a)R_(8b), —X₁NR_(8a)S(O)₀₋₂R_(8b); wherein each X₁is independently C₁₋₄alkylene; and X₁ optionally has 1 to 3 hydrogensreplaced with a group selected from hydroxy, halo, cyano, C₁₋₄alkyl,halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy and halo-substituted-C₁₋₄alkoxy;X₂ is selected from a bond and C₁₋₄alkylene; wherein R_(8a) and R_(8b)are independently selected from hydrogen, C₁₋₆alkyl,halo-substituted-C₁₋₆alkyl, C₃₋₈cycloalkyl, heteroaryl andC₃₋₈heterocycloalkyl; wherein the cycloalkyl, heterocycloalkyl orheteroaryl of R_(8a) or R_(8b) is optionally substituted with 1 to 3radicals independently selected from amino, cyano, C₁₋₄alkyl,C₁₋₄alkoxy, halo-substituted-C₁₋₄alkyl and halo-substituted-C₁₋₄alkoxy;with the proviso that R_(8b) is not hydrogen when R₁ is selected from—X₁NHC(O)OR_(8b) and —X₁NR_(8a)S(O)₀₋₂R_(8b);

R₂, R₃, R₅ and R₆ are independently selected from hydrogen, halo, cyano,C₁₋₄alkyl, halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy andhalo-substituted-C₁₋₄alkoxy; with the proviso that when R₅ is fluoro andR₁ is selected from hydrogen, —X₁R_(8a), —X₁OX₂R_(8a),—X₁C(O)NR_(8a)R_(8b), —X₁NR_(8a)X₂R_(8b), —X₁NR_(8a)C(O)X₂OR_(8b) and—X₁NR_(8a)S(O)₀₋₂R_(8b), R₃ and R₆ are not both hydrogen;

R₄ is selected from —R₉ and —NR₁₀R₁₁; wherein R₉ is selected fromC₁₋₆alkyl, C₃₋₈cycloalkyl, C₃₋₈heterocycloalkyl, aryl and heteroaryl;wherein said alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl ofR₉ is optionally substituted with 1 to 3 radicals independently selectedfrom halo, cyano, halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy andhalo-substituted-C₁₋₄alkoxy; and R₁₀ and R₁₁ are independently selectedfrom hydrogen and R₉;

R₇ is selected from hydrogen, C₁₋₄alkyl, C₃₋₅cycloalkyl andC₃₋₅heterocycloalkyl; wherein said alkyl, cycloalkyl or heterocycloalkylof R₇ is optionally substituted with 1 to 3 radicals independentlyselected from halo, cyano, hydroxyl, C₁₋₄alkyl,halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy and halo-substituted-C₁₋₄alkoxy;and the N-oxide derivatives, prodrug derivatives, protected derivatives,the tautomers, individual isomers and mixture of isomers thereof; andthe pharmaceutically acceptable salts and solvates (e.g. hydrates) ofsuch compounds.

In a second aspect, the present invention provides a pharmaceuticalcomposition which contains a compound of Formula I or a N-oxidederivative, individual isomers and mixture of isomers thereof; or apharmaceutically acceptable salt thereof, in admixture with one or moresuitable excipients.

In a third aspect, the present invention provides a method of treating adisease in an animal in which inhibition of kinase activity,particularly B-Raf activity, can prevent, inhibit or ameliorate thepathology and/or symptomology of the diseases, which method comprisesadministering to the animal a therapeutically effective amount of acompound of Formula I or a N-oxide derivative, individual isomers andmixture of isomers thereof, or a pharmaceutically acceptable saltthereof.

In a fourth aspect, the present invention provides the use of a compoundof Formula I in the manufacture of a medicament for treating a diseasein an animal in which kinase activity, particularly B-Raf activity,particularly mutant B-raf (for example V600E), contributes to thepathology and/or symptomology of the disease.

In a fifth aspect, the present invention provides a process forpreparing compounds of Formula I and the N-oxide derivatives, prodrugderivatives, protected derivatives, individual isomers and mixture ofisomers thereof, and the pharmaceutically acceptable salts thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: FIG. 1 illustrates that the addition of a MEK small moleculeinhibitor can reverse the induced ERK signaling, cell growth andtransformation caused by a Raf small molecule inhibitor.

DETAILED DESCRIPTION OF THE INVENTION Definitions

“Alkyl” as a group and as a structural element of other groups, forexample halo-substituted-alkyl and alkoxy, can be eitherstraight-chained or branched. C₁₋₄-alkoxy includes, methoxy, ethoxy, andthe like. Halo-substituted-C₁₋₄alkyl means and alkyl group (branched orunbranched) wherein any of the hydrogens can be substituted with ahalogen. For example, halo-substituted-C₁₋₄alkyl can be trifluoromethyl,difluoroethyl, pentafluoroethyl, and the like. Similarly,hydroxy-substituted-C₁₋₆alkyl means and alkyl group (branched orunbranched) wherein any of the hydrogens can be substituted with ahydroxyl. For example, hydroxy-substituted-C₁₋₆alkyl includes2-hydroxyethyl, and the like. Similarly, cyano-substituted-C₁₋₆alkylmeans and alkyl group (branched or unbranched) wherein any of thehydrogens can be substituted with cyano.

“Aryl” means a monocyclic or fused bicyclic aromatic ring assemblycontaining six to ten ring carbon atoms. For example, aryl may be phenylor naphthyl, preferably phenyl. “Arylene” means a divalent radicalderived from an aryl group.

“Cycloalkyl” means a saturated or partially unsaturated, monocyclic,fused bicyclic or bridged polycyclic ring assembly containing the numberof ring atoms indicated. For example, C₃₋₁₀cycloalkyl includescyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

“Heteroaryl” is as defined for aryl above where one or more of the ringmembers is a heteroatom. For example, heteroaryl includes pyridyl,indolyl, indazolyl, quinoxalinyl, quinolinyl, benzofuranyl,benzopyranyl, benzothiopyranyl, benzo[1,3]dioxole, imidazolyl,benzo-imidazolyl, pyrimidinyl, furanyl, oxazolyl, isoxazolyl, triazolyl,tetrazolyl, pyrazolyl, thienyl, etc.

“Heterocycloalkyl” means cycloalkyl, as defined in this application,provided that one or more of the ring carbons indicated, are replaced bya moiety selected from —O—, —N═, —NR—, —C(O)—, —S—, —S(O)— or —S(O)₂—,wherein R is hydrogen, C₁₋₄alkyl or a nitrogen protecting group. Forexample, C₃₋₈heterocycloalkyl as used in this application to describecompounds of the invention includes 2H-pyran, 4H-pyran, piperidine,1,4-dioxane, morpholine, 1,4-dithiane, thiomorpholino,imidazolidin-2-one, tetrahydrofuran, piperazine, 1,3,5-trithiane,pyrrolidine, pyrrolidinyl-2-one, piperidine, piperidinone,1,4-dioxa-8-aza-spiro[4.5]dec-8-yl, etc.

“Halogen” (or halo) represents chloro, fluoro, bromo or iodo.

“pMEK” means phosphorylated Mek.

“pERK” means phosphorylated ERK.

“Treat”, “treating” and “treatment” refer to a method of alleviating orabating a disease and/or its attendant symptoms.

Compounds of the invention are named using Chemdraw Ultra (Version 10.0)and/or ChemAxon Name Generator (JChem Version 5.3.1.0).

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention provides compounds, compositions and methods forthe treatment of kinase related disease, particularly B-Raf kinaserelated diseases; for example, metastatic melanomas, solid tumors, braintumors such as Glioblastoma multiform (GBM), acute myelogenous leukemia(AML), prostate cancer, gastric cancer, papillary thyroid carcinoma,ovarian low-grade carcinoma, and colorectal cancer.

In one embodiment, with reference to compounds of Formula I, R₁ isselected from —X₁R_(8a) and —X₁NHC(O)OR_(8b); wherein each X₁ isindependently C₁₋₄alkylene; and X₁ optionally has 1 to 3 hydrogensreplaced with a group selected from hydroxy, halo, cyano, C₁₋₄alkyl andhalo-substituted-C₁₋₄alkyl; wherein R_(8a) and R_(8b) are independentlyselected from hydrogen and C₁₋₆alkyl; with the proviso that R_(8b) isnot hydrogen when R₁ is —X₁NHC(O)OR_(8b);

In another embodiment are compounds of Formula Ia:

in which: Y is selected from N and CR₆; R₂, R₃, R₅ and R₆ areindependently is selected from hydrogen, halo, cyano, C₁₋₄alkyl,halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy and halo-substituted-C₁₋₄alkoxy;with the proviso that when R₅ is fluoro and R₁ is selected fromhydrogen, —X₁R_(8a), —X₁OX₂R_(8a), —X₁C(O)NR_(8a)R_(8b),—X₁NR_(8a)X₂R_(8b), —X₁NR_(8a)C(O)X₂OR_(8b) and —X₁NR_(8a)S(O)₀₋₂R_(8b),R₃ and R₆ are not both hydrogen; R₄ is selected from —R₉ and —NR₁₀R₁₁;wherein R₉ is selected from C₁₋₆alkyl, C₃₋₈cycloalkyl,C₃₋₈heterocycloalkyl, aryl and heteroaryl; wherein said alkyl,cycloalkyl, heterocycloalkyl, aryl or heteroaryl of R₉ is optionallysubstituted with 1 to 3 radicals independently selected from halo,cyano, C₁₋₄ alkyl, halo-substituted-C₁₋₄ alkyl, C₁₋₄ alkoxy andhalo-substituted-C₁₋₄alkoxy; and R₁₀ and R₁₁ are independently selectedfrom hydrogen and R₉; and R₇ is selected from hydrogen, C₁₋₄alkyl,C₃₋₅cycloalkyl and C₃₋₅heterocycloalkyl; wherein said alkyl, cycloalkylor heterocycloalkyl of R₇ is optionally substituted with 1 to 3 radicalsindependently selected from halo, cyano, hydroxyl, C₁₋₄alkyl,halo-substituted-C₁₋₄alkyl, C₁₋₄ alkoxy and halo-substituted-C₁₋₄alkoxy.

In a further embodiment, R₄ is —R₉; wherein R₉ is selected fromC₁₋₃alkyl and C₃₋₈cycloalkyl; wherein said alkyl or cycloalkyl of R₉ isoptionally substituted with 1 to 3 radicals independently selected fromhalo and halo-substituted-C₁₋₄alkyl.

In a further embodiment, R₂ is selected from hydrogen and fluoro; R₃ isselected from chloro, fluoro and methyl; R₅ is selected hydrogen, fromchloro and fluoro; Y is selected from N and CR₆; and R₆ is selected fromhydrogen and fluoro.

In a further embodiment are compounds selected from: methylN-[(2S)-1-({4-[3-(3-chloro-5-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-[(4-{3-[2-fluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-[(4-{3-[3-chloro-5-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2,6-difluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-[(4-{3-[2,6-difluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate;methylN-[(2S)-1-{[4-(3-{2-fluoro-3-[(3,3,3-trifluoropropane)sulfonamido]phenyl}-1-(propan-2-yl)-1H-pyrazol-4-yl)pyrimidin-2-yl]amino}propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(3-chloro-2-methanesulfonamidopyridin-4-yl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(3-fluoro-2-methanesulfonamidopyridin-4-yl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2-chloro-3-ethanesulfonamido-4,5-difluorophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2,4-difluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[1-(propan-2-yl)-3-(2,4,5-trifluoro-3-methanesulfonamidophenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate; methylN-[(2S)-1-({4-[3-(3-ethanesulfonamido-2,4-difluorophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-2-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propyl]carbamate;methylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(oxan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-[(4-{3-[2,4-difluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(3-cyclopropanesulfonamido-2,5-difluorophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(5-chloro-3-cyclopropanesulfonamido-2-fluorophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;and methylN-[(2S)-1-[(4-{3-[5-chloro-2-fluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate.

In another embodiment are compounds of Formula Ib:

in which: R₃ is selected from chloro, fluoro and methyl; R₅ is selectedfrom fluoro and chloro; and R₇ is selected from ethyl and isopropyl.

In a further embodiment are compounds selected from: methylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2,5-difluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-ethyl-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2-fluoro-3-methanesulfonamido-5-methylphenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2-chloro-3-methanesulfonamido-5-methylphenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2-chloro-5-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2R)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1II-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;methylN-[(2S)-1-({4-[3-(2,5-dichloro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate;and methylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate.

In another embodiment are compounds selected from:N-[5-chloro-3-(4-{2-[(2-cyanoethyl)amino]pyrimidin-4-yl}-1-(propan-2-yl)-1H-pyrazol-3-yl)-2-fluorophenyl]methanesulfonamide;N-{5-chloro-3-[4-(2-{[2-(dimethylamino)ethyl]amino}pyrimidin-4-yl)-1-(propan-2-yl)-1H-pyrazol-3-yl]-2-fluorophenyl}methanesulfonamide;N-(5-chloro-2-fluoro-3-{4-[2-(methylamino)pyrimidin-4-yl]-1-(propan-2-yl)-1H-pyrazol-3-yl}phenyl)methanesulfonamide;andN-{3-[4-(2-aminopyrimidin-4-yl)-1-(propan-2-yl)-1H-pyrazol-3-yl]-5-chloro-2-fluorophenyl}methanesulfonamide.

In a further embodiment are compounds selected from the Examples andTables, infra.

In a further embodiment are intermediate compounds selected from:3-Bromo-5-chloro-2-fluoroaniline;cyano-(2-methylthio-pyrimidin-4-yl)-acetic acid tert-butyl ester;1-Isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine;2-((2-Benzylidene-1-ethylhydrazinyl)methylene)-malononitrile;1-(3-Amino-1-isopropyl-1H-pyrazol-4-yl)ethanone;1-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)ethanone;1-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)ethanone;1-(3-Iodo-1-methyl-1H-pyrazol-4-yl)ethanone;3-(Dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)prop-2-en-1-one;3-(Dimethylamino)-1-(3-iodo-1-ethyl-1H-pyrazol-4-yl)prop-2-en-1-one;3-(Dimethylamino)-1-(3-iodo-1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one;4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-amine;4-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidin-2-amine;4-(3-Iodo-1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine;4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ol;2-Chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine; (S)-Methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate;(R)-Methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate;(S)-tert-butyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate;3-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propanenitrile;4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-N-methylpyrimidin-2-amine;N¹-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-yl)-N²,N²-dimethylethane-1,2-diamine;N-(3-bromo-2,4-difluorophenyl)propane-1-sulfonamide;3-Fluoro-4-iodopyridin-2-amine; 3-chloro-4-iodopyridin-2-amine;3-Bromo-2,5,6-trifluoroaniline; 2,4-Dibromo-3,6-dichloroaniline;3-bromo-2-chloro-5-methylaniline; 3-bromo-2,5-difluoroaniline;3-Bromo-5-chloro-2-fluorobenzoic acid; Tert-butyl3-bromo-5-chloro-2-fluorophenylcarbamate; tert-butyl3-bromo-2-fluoro-5-methylphenylcarbamate; Tert-butyl5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate;tert-butyl2,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate;N-(2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propane-1-sulfonamide;2-(2-fluoro-3-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane;2,5-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;2-chloro-5-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;2,5-dichloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;2-chloro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;tert-butyl2-fluoro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate;3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine;2,3,6-trifluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine;3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline; and3-methoxy-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline.

The present invention also includes all suitable isotopic variations ofthe compounds of the invention, or pharmaceutically acceptable saltsthereof. An isotopic variation of a compound of the invention or apharmaceutically acceptable salt thereof is defined as one in which atleast one atom is replaced by an atom having the same atomic number butan atomic mass different from the atomic mass usually found in nature.

Examples of isotopes that may be incorporated into the compounds of theinvention and pharmaceutically acceptable salts thereof include but arenot limited to isotopes of hydrogen, carbon, nitrogen and oxygen such as²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ³⁵S, ¹⁸F, ³⁶Cl and ¹²³I. Certainisotopic variations of the compounds of the invention andpharmaceutically acceptable salts thereof, for example, those in which aradioactive isotope such as ³H or ¹⁴C is incorporated, are useful indrug and/or substrate tissue distribution studies. In particularexamples, ³H and ¹⁴C isotopes may be used for their ease of preparationand detectability. In other examples, substitution with isotopes such as²H may afford certain therapeutic advantages resulting from greatermetabolic stability, such as increased in vivo half-life or reduceddosage requirements. Isotopic variations of the compounds of theinvention or pharmaceutically acceptable salts thereof can generally beprepared by conventional procedures using appropriate isotopicvariations of suitable reagents.

Some Raf inhibitors, in addition to increasing MEK and ERK signaling inwild-type B-Raf cells, also induce cell growth in cancer cell lines andcause transformation and growth in fibroblasts. The induction ofdownstream signaling has previously been attributed to published Rafpathway feedback loops. However, induction of pMEK and pERK can occurwithin minutes of Raf inhibitor treatment, even before reported feedbackphosphorylation events are seen on B-Raf and C-Raf. The induction ofsignaling and cell growth both occur in a biphasic pattern, with lowcompound concentrations (0.01-0.1 μM) causing maximal induction, andhigher compound concentrations (1-10 μM) causing less profoundinduction. Such a biphasic pattern is also observed in biochemicalassays with purified wild-type B-Raf or C-Raf and is suggestive of amechanism involving the interaction of two signaling subunits. Inaddition, Raf dimerization can up regulate pMEK, not throughtrans-phosphorylation of Raf molecules but presumably by aconformational activation of the kinase. Consistent with that model, Rafinhibitor treatment induces B-Raf/C-Raf dimer formation in cells. Inaddition, knockdown of A- or B-Raf with siRNA does not abrogate the Rafinhibitor induction of pMEK and pERK, and knockdown of C-Raf onlyslightly decreases the induction. Notably, knockdown of K-Ras in K-Rasmutant cells also only slightly decreases the induction, implying thatthis effect is not primarily mediated by Ras. Taken together, the datasuggest a model in which inhibitor binding to one Raf molecule inducesdimerization and conformational activation of a partner Raf molecule inthe dimer. This can explain why wild-type Raf and mutant Ras tumors areinsensitive to selective Raf kinase inhibitors and might also haveimportant implications for toxicity, since induction of strong mitogenicsignaling could lead to hyper proliferation of normal tissues.Understanding the Raf inhibitor induction mechanism may lead to thedesign of improved inhibitors.

The addition of a MEK inhibitor in combination with a Raf inhibitorleads to a significant inhibition of ERK signaling and consequently adecrease in cellular proliferation and transformation. Since MEKinhibitor treatments alone have led to dose limiting toxicities in theclinic, a Raf plus MEK inhibitor combination may represent a superiortreatment strategy.

The present invention also includes combinations of the BRaf inhibitorsdisclosed in this invention with other agents. In particular, thepresent invention provides for combinations with MEK1/2 inhibitors. FIG.1 illustrates that the addition of a MEK small molecule inhibitor canreverse the induced ERK signaling, cell growth and transformation causedby a Raf small molecule inhibitor. For example, a compound of Formula I(compound 9 of the invention, namely: ((S)-methyl1-(4-(3-(5-chloro-2-fluoro-3-(methylsulfonamido)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate)can lead to an induction in cellular proliferation, seen as a negativeinhibition in FIG. 1 of a Cell Titer Glo assay using SW620 cells. TheY-axis shows negative and positive inhibition. Each experiment is shownas a series of 9 serial dilutions between 10 and 0.002 μM. Compound A1(N-(4-methyl-3-(1-(6-(4-methylpiperazin-1-ylamino)pyrimidin-4-yl)-1H-imidazol-2-ylamino)phenyl)-3-(trifluoromethyl)benzamide)is a control, A3 is a MEK inhibitor (PD0325901). Compound 9 is tested inthe absence and presence of 1 μM, 0.1 μM and 0.01 μM of MEK inhibitorA3.

Pharmacology and Utility

Compounds of the invention modulate the activity of kinases and, assuch, are useful for treating diseases or disorders in which kinasescontribute to the pathology and/or symptomology of the disease. Examplesof kinases that are inhibited by the compounds and compositionsdescribed herein and against which the methods described herein areuseful include, but are not limited to, B-Raf, including mutant forms ofB-Raf.

The mitogen-activated protein kinase (MAPK) pathway mediates theactivity of a number of effector molecules which coordinate to controlcellular proliferation, survival, differentiation and migration.Stimulation of cells by, for example, growth factors, cytokines orhormones results in the plasma membrane-associated Ras becomingGTP-bound and thereby activated to recruit Raf. This interaction inducesthe kinase activity of Raf leading to direct phosphorylation of MAPK/ERK(MEK), which in turn phosphorylates the extracellular signal-relatedkinase (ERK). Activated ERK then phosphorylates a wide array of effectormolecules, for example, kinases, phosphatases, transcription factors andcytoskeletal proteins. Therefore, the Ras-Raf-MEK-ERK signaling pathwaytransmits signals from cell surface receptors to the nucleus and isessential, for example, in cell proliferation and survival. Theregulation of this signaling cascade is further enriched by the multipleisoforms of Ras (including K-Ras, N-Ras and H-Ras), Raf (A-Raf, B-Raf,C-Raf/Raf-1), MEK (MEK-1 and MEK-2) and ERK (ERK-1 and ERK-2). Since10-20% of human cancers harbor oncogenic Ras mutations and many humancancers have activated growth factor receptors, this pathway is an idealtarget for intervention.

The essential role and the position of Raf in many signaling pathwayshas been demonstrated from studies using deregulated and dominantinhibitory Raf mutants in mammalian cells as well as from studiesemploying biochemical and genetic techniques to model organisms. In thepast, the focus on Raf being an anti-tumor drug target centered on itsfunction as a downstream effector of Ras. However, recent findingssuggest that Raf may have a prominent role in the formation of certaintumors with no requirement of an oncogenic Ras allele. In particular,activating alleles of B-Raf and N-Ras have been identified in ˜70% ofmelanomas, 40% of papillary thyroid carcinoma, 30% of ovarian low-gradecarcinoma, and 10% of colorectal cancers. Mutations in K-Ras occur inapproximately 90% of pancreatic cancers. Most B-Raf mutations are foundwithin the kinase domain, with a single substitution (V600E) accountingfor at least 80%. The mutated B-Raf proteins activate the Raf-MEK-ERKpathway either via elevated kinase activity towards MEK or viaactivating C-Raf.

Therefore, development of a kinase inhibitor for B-Raf provides a newtherapeutic opportunity for treatment of many types of human cancers,especially for metastatic melanomas, solid tumors, brain tumors such asGlioblastoma multiform (GBM), acute myelogenous leukemia (AML), lungcancer; papillary thyroid carcinoma, ovarian low-grade carcinoma, andcolorectal cancer. Several Raf kinase inhibitors have been described asexhibiting efficacy in inhibiting tumor cell proliferation in vitroand/or in vivo assays (see, for example, U.S. Pat. Nos. 6,391,636,6,358,932, 6,037,136, 5,717,100, 6,458,813, 6,204,467, and 6,268,391).Other patents and patent applications suggest the use of Raf kinaseinhibitors for treating leukemia (see, for example, U.S. Pat. Nos.6,268,391, 6,204,467, 6,756,410, and 6,281,193; and abandoned U.S.Patent Application Nos. 20020137774 and 20010006975), or for treatingbreast cancer (see, for example, U.S. Pat. Nos. 6,358,932, 5,717,100,6,458,813, 6,268,391, 6,204,467 and 6,911,446). Data demonstrates thatRaf kinase inhibitors can significantly inhibit signaling through theMAPK pathway, leading to dramatic shrinkage in B-Raf (V600E) tumors.

The compounds of the present invention inhibit cellular processesinvolving B-Raf kinase by blocking the signal cascade in these cancercells and ultimately inducing stasis and/or death of the cells.

In accordance with the foregoing, the present invention further providesa method for preventing or treating lung carcinoma, prostate cancer,gastric cancer, pancreatic carcinoma, bladder carcinoma, coloncarcinoma, myeloid disorders, prostate cancer, thyroid cancer, melanoma,adenomas and carcinomas of the ovary, eye, liver, biliary tract, andnervous system. Further, the present invention further provides a methodfor preventing or treating any of the diseases or disorders describedabove in a subject in need of such treatment, which method comprisesadministering to said subject a therapeutically effective amount (See,“Administration and Pharmaceutical Compositions”, infra) of a compoundof Formula I or a pharmaceutically acceptable salt thereof. For any ofthe above uses, the required dosage will vary depending on the mode ofadministration, the particular condition to be treated and the effectdesired.

Administration and Pharmaceutical Compositions

In general, compounds of the invention will be administered intherapeutically effective amounts via any of the usual and acceptablemodes known in the art, either singly or in combination with one or moretherapeutic agents. A therapeutically effective amount may vary widelydepending on the severity of the disease, the age and relative health ofthe subject, the potency of the compound used and other factors. Ingeneral, satisfactory results are indicated to be obtained systemicallyat daily dosages of from about 0.03 to 30 mg/kg per body weight. Anindicated daily dosage in the larger mammal, e.g. humans, is in therange from about 0.5 mg to about 2000 mg, conveniently administered,e.g. in divided doses up to four times a day or in retard form. Suitableunit dosage forms for oral administration comprise from ca. 1 to 500 mgactive ingredient.

Compounds of the invention can be administered as pharmaceuticalcompositions by any conventional route, in particular enterally, e.g.,orally, e.g., in the form of tablets or capsules, or parenterally, e.g.,in the form of injectable solutions or suspensions, topically, e.g., inthe form of lotions, gels, ointments or creams, or in a nasal orsuppository form. Pharmaceutical compositions comprising a compound ofthe present invention in free form or in a pharmaceutically acceptablesalt form in association with at least one pharmaceutically acceptablecarrier or diluent can be manufactured in a conventional manner bymixing, granulating or coating methods. For example, oral compositionscan be tablets or gelatin capsules comprising the active ingredienttogether with a) diluents, e.g., lactose, dextrose, sucrose, mannitol,sorbitol, cellulose and/or glycine; b) lubricants, e.g., silica, talcum,stearic acid, its magnesium or calcium salt and/or polyethyleneglycol;for tablets also c) binders, e.g., magnesium aluminum silicate, starchpaste, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose and or polyvinylpyrrolidone; if desired d)disintegrants, e.g., starches, agar, alginic acid or its sodium salt, oreffervescent mixtures; and/or e) absorbents, colorants, flavors andsweeteners. Injectable compositions can be aqueous isotonic solutions orsuspensions, and suppositories can be prepared from fatty emulsions orsuspensions. The compositions may be sterilized and/or containadjuvants, such as preserving, stabilizing, wetting or emulsifyingagents, solution promoters, salts for regulating the osmotic pressureand/or buffers. In addition, they may also contain other therapeuticallyvaluable substances. For example, compounds of the invention can beformulated into a microemulsion pre concentrate (MEPC). Compounds ofFormula I can be prepared at 40 mg/ml in a mixture of 56% PEG400, 29%cremophor EL and 15% oleic acid. The mixture, without a compound ofFormula I, is first prepared by vortexing/shaking. A compound of theinvention is added and sonication is used to disperse the powder intothe vehicle. The mixture is heated to 80° C. in a water bath for aboutan hour stirring, with sonication, every 15 minutes. This mixture isphysically and chemically stable at room temperature for about one week.

Suitable formulations for transdermal applications include an effectiveamount of a compound of the present invention with a carrier. A carriercan include absorbable pharmacologically acceptable solvents to assistpassage through the skin of the host. For example, transdermal devicesare in the form of a bandage comprising a backing member, a reservoircontaining the compound optionally with carriers, optionally a ratecontrolling barrier to deliver the compound to the skin of the host at acontrolled and predetermined rate over a prolonged period of time, andmeans to secure the device to the skin. Matrix transdermal formulationsmay also be used. Suitable formulations for topical application, e.g.,to the skin and eyes, are preferably aqueous solutions, ointments,creams or gels well-known in the art. Such may contain solubilizers,stabilizers, tonicity enhancing agents, buffers and preservatives.

Compounds of the invention can be administered in therapeuticallyeffective amounts in combination with one or more therapeutic agents(pharmaceutical combinations). For example, synergistic effects canoccur with other anti-tumor or anti-proliferative agents, for example,mitotic inhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, antibodies,cytotoxics, anti-hormones, anti-androgens, an anti-angiogenesis agent,kinase inhibitor, pan kinase inhibitor or growth factor inhibitor.

Compounds of the invention can be administered in therapeuticallyeffective amounts in combination with one or more suitable excipientsselected from corn starch, potato starch, tapioca starch, starch paste,pre-gelatinized starch, sugars, gelatin, natural gums, synthetic gums,sodium alginate, alginic acid, tragacanth, guar gum, cellulose, ethylcellulose, cellulose acetate, carboxymethyl cellulose calcium, sodiumcarboxymethylcellulose, methyl cellulose, hydroxypropyl methylcellulose,microcrystalline cellulose, magnesium aluminum silicate, polyvinylpyrrolidone, talc, calcium carbonate, powdered cellulose, dextrates,kaolin, mannitol, silicic acid, sorbitol, agar-agar, sodium carbonate,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, clays, sodium stearate, calcium stearate, magnesium stearate,stearic acid, mineral oil, light mineral oil, glycerin, sorbitol,mannitol, polyethylene glycol, other glycols, sodium lauryl sulfate,hydrogenated vegetable oil, peanut oil, cottonseed oil, sunflower oil,sesame oil, olive oil, corn oil, soybean oil, zinc stearate, sodiumoleate, ethyl oleate, ethyl laureate, silica, and combinations thereof.

An embodiment of the invention is a method of claim 12 or 13, furthercomprising administering to the subject an additional therapeutic agent.The additional therapeutic agent comprises an anticancer drug, a painmedication, an antiemetic, an antidepressant or an anti-inflammatoryagent. Further, the additional therapeutic agent is a different Rafkinase inhibitor or an inhibitor of MEK, mTOR, HSP90, AKT, PI3K, CDK9,PAK, Protein Kinase C, a MAP kinase, a MAPK Kinase, or ERK and isadministered to the subject concurrently with a compound of theinvention.

For example, the addition of a MEK inhibitor in combination with a Rafinhibitor leads to a significant inhibition of ERK signaling andconsequently a decrease in cellular proliferation and transformation.Since MEK inhibitor treatments alone have led to dose limitingtoxicities in the clinic, a Raf plus MEK inhibitor combinationrepresents a superior treatment strategy. Examples of MEK inhibitors areAS703026 (EMD Serono); MSC1936369B (EMD Serono); GSK1120212(GlaxoSmithKline); AZD6244 (Memorial Sloan-Kettering Cancer Center);PD-0325901 (Pfizer); ARRY-438162 (Array BioPharma); RDEA 119 (ArdeaBiosciences, Inc.); GDC0941 (Genentech); GDC0973 (Genentech); TAK-733(Millennium Pharmaceuticals, Inc.); RO5126766 (Hoffmann-La Roche); andXL-518 (Exelixis).

In another embodiment of the invention are combinations and methods oftreating cancer comprising a therapeutically effective amount of acompound of the Summary of the Invention (Raf inhibitor) and at leastone MEK protein kinase inhibitor.

Where the compounds of the invention are administered in conjunctionwith other therapies, dosages of the co-administered compounds will ofcourse vary depending on the type of co-drug employed, on the specificdrug employed, on the condition being treated and so forth.

The invention also provides for a pharmaceutical combinations, e.g. akit, comprising a) a first agent which is a compound of the invention asdisclosed herein, in free form or in pharmaceutically acceptable saltform, and b) at least one co-agent. The kit can comprise instructionsfor its administration.

The terms “co-administration” or “combined administration” or the likeas utilized herein are meant to encompass administration of the selectedtherapeutic agents to a single patient, and are intended to includetreatment regimens in which the agents are not necessarily administeredby the same route of administration or at the same time.

The term “pharmaceutical combination” as used herein means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. a compound of Formula I and a co-agent, are bothadministered to a patient simultaneously in the form of a single entityor dosage. The term “non-fixed combination” means that the activeingredients, e.g. a compound of Formula I and a co-agent, are bothadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific time limits, wherein suchadministration provides therapeutically effective levels of the 2compounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of 3 or more activeingredients.

Processes for Making Compounds of the Invention

The present invention also includes processes for the preparation ofcompounds of the invention. In the reactions described, it can benecessary to protect reactive functional groups, for example hydroxy,amino, imino, thio or carboxy groups, where these are desired in thefinal product, to avoid their unwanted participation in the reactions.Conventional protecting groups can be used in accordance with standardpractice, for example, see T. W. Greene and P. G. M. Wuts in “ProtectiveGroups in Organic Chemistry”, John Wiley and Sons, 1991.

Compounds of Formula I can be prepared by proceeding as in the followingReaction Scheme I:

in which R₁, R₂, R₃, R₄, R₅, R₇, and Y are as defined in the Summary ofthe Invention, P is a suitable protecting group (for example, MEM, MOM,SEM, R₄SO₂, and the like), and M is a leaving group (for example,chloro, bromo, iodo, methanesulfonyloxy, p-toluenesulfonyloxy, and thelike). A compound of Formula I can be synthesized by removal ofprotecting group P from a compound of Formula 2 (for example, bytreatment with a strong acid such as hydrogen chloride in the presenceof a protic solvent such as methanol or water when P is MEM, MOM, orSEM; or by treatment with aqueous or methanolic sodium or potassiumcarbonate, optionally in the presence of a cosolvent such as toluene,when P is a second sulfonyl group R₄SO₂).

Compounds of Formula 2 can be prepared by reacting a compound of Formula3 with an alkylating agent of Formula 4 in the presence of a suitablesolvent (for example, DMF, DMSO, and the like) and a suitable base (forexample, potassium carbonate, sodium hydride, and the like). Thereaction proceeds in a temperature range of about 0° C. to about 150° C.and can take up to about 24 hours to complete. The reaction mixture isoptionally further reacted to remove any protecting groups.

Compounds of Formula I can also be prepared by proceeding as in thefollowing Reaction Scheme II:

in which R₁, R₂, R₃, R₄, R₅, and Y are as defined in the Summary of theInvention. A compound of Formula I can be synthesized by reacting acompound of Formula 5 with a sulfonylating reagent of Formula 6 in thepresence of a suitable base (for example, pyridine, triethylamine,4-(N,N-dimethylamino)pyridine, and the like) and a suitable solvent(such as pyridine, dichloromethane, 2-methylTHF, and the like). Thereaction proceeds in a temperature range of about 0° C. to about 100° C.and can take up to about 24 hours to complete. The reaction mixture isoptionally further reacted to remove any protecting groups. In someinstances, the sulfonylating reagent can react twice to produce abis-sulfonyl derivative. In this instance, the bis-sulfonyl compound canbe converted to a compound of Formula a by treatment with a suitablebase (for example, sodium or potassium hydroxide, or sodium or potassiumcarbonate) in the presence of a protic solvent such as methanol orwater, optionally in the presence of a cosolvent such as toluene or2-methylTHF. The reaction takes place in a temperature range of about20° C. to about 100° C. and can take up to about 24 hours to complete.

Compounds of Formula I can also be prepared by proceeding as in thefollowing Reaction Scheme III:

in which R₁, R₂, R₃, R₄, R₅, and Y are as defined in the Summary of theInvention, R₅₀ is a leaving group (for example, iodo, bromo, chloro,trifluoromethanesulfonyloxy, and the like), each R′ can be, for examplehydrogen, methyl, and the like, or the two R′ groups can be joinedtogether to form a cyclic boronate ester. The two R₉₀ groups can each behydrogen, or the two R₉₀ groups, taken together, can represent asuitable nitrogen protecting group (for example, one R₉₀ can be hydrogenand the other can be BOC). A compound of Formula Ia can be synthesizedby reacting a compound of Formula 7 with a compound of Formula 8 in thepresence of a suitable transition metal catalyst (for example,tetrakis(triphenylphosphinepalladium)(0) or PdCl₂(dppf), a suitablesolvent (for example, DME, dioxane, toluene, ethanol, and the like) anda suitable base (for example, anhydrous potassium carbonate or aqueoussodium carbonate solution, and the like). The reaction proceeds in atemperature range of about 20° C. to about 120° C. and can take up toabout 24 hours to complete. The reaction mixture is optionally furtherreacted to remove any protecting groups.

A compound of Formula I can also be prepared by a similar Suzukireaction protocol in which the a compound of Formula 7 is reacted with acompound of Formula 8a to generate a compound of Formula 9. Followingdeprotection of the R₉₀ groups, a sulfonylation reaction, as describedfor Reaction Scheme II, generates a compound of Formula Ia.

It will be appreciated by one skilled in the art that otherorganometallic coupling reactions, for example using tin reagents(Stille coupling) or zinc reagents (Negishi coupling), might also beemployed in place of the Suzuki coupling reaction using boron reagentsdescribed in Reaction Scheme III.

Compounds of Formula 7 can be prepared by proceeding as in the followingReaction Scheme IV:

In this instance, M is a leaving group (for example, chloro, bromo,iodo, methanesulfonyl, and the like), R₅₀ is a leaving group (forexample, iodo, bromo, chloro, trifluoromethanesulfonyloxy, and thelike), and R₇ is as defined in the Summary of the Invention. A compoundof Formula 7 can be prepared by reacting an amine compound of Formula 10with a compound of Formula 11. The reaction is performed in the presenceof a suitable base (for example, triethylamine, potassium carbonate, andthe like) in a solvent such as isopropanol, DMSO, NMP, or dioxane at atemperature from about 25 to about 120° C. In some instances furthertransformations of the newly-introduced R₁ group may subsequently beperformed to arrive at the final intended R₁ group.

Compounds of Formula 11a, which are a subset of compounds of Formula 11in which M is methanesulfonyl, can be prepared by proceeding as in thefollowing Reaction Scheme V:

In which R₇ and R₅₀ are as defined in the Summary of the Invention. Acompound of Formula 11a can be prepared by reacting a compound ofFormula 12 with a suitable oxidizing system (for example,m-chloroperbenzoic acid in a solvent of dichloromethane, or Oxone™ inaqueous methanol, and the like), at a temperature of about −78° C. toabout 50° C. The reaction takes up to about 24 hours to complete.

A compound of Formula 12 in which R₅₀ is chloro, bromo, or iodo can inturn be prepared by reacting an amine compound of Formula 13 with asuitable diazotization reagent system (for example, nitrous acid inconjunction with a copper (I) halide salt, isoamyl nitrite inconjunction with copper (I) iodide/methylene iodide, isoamyl nitrite inconjunction with boron trifluoride, iodine, and potassium iodide inacetonitrile, and the like). The reaction takes place at a temperatureof about 0 to about 80° C., and takes from about 1 to about 6 hours tocomplete.

A compound of Formula 13 can be prepared by reacting a compound ofFormula 14 with a compound of Formula 3, as described for ReactionScheme I.

A compound of Formula 14 can be prepared by cyclization of anenaminonitrile compound of Formula 15 with hydrazine or a hydrazine saltin a suitable solvent (for example, ethanol and the like). The reactiontakes place at a temperature of about 25 to about 100° C., and can takefrom about 1 hour to about 24 hours to complete.

Alternatively, a compound of Formula 13 can be prepared from a compoundof Formula 15 in a one-step procedure by reacting a compound of Formula15 with a monosubstituted hydrazine R₇NH—NH₂. The reaction takes placeat a temperature of about 25 to about 100° C., and can take from about 1hour to about 24 hours to complete.

A compound of Formula 15 can in turn be prepared by reaction of acompound of Formula 16 with DMF DMA or Bredereck's reagent, optionallyin the presence of a co-solvent such as DMF, at a temperature of about50 to about 150° C. The reaction takes about 1 to about 24 hours tocomplete.

A compound of Formula 16 can be prepared by treatment of a compound ofFormula 17 with a suitable acid (for example, p-toluenesulfonic acid,and the like) in a suitable inert solvent (for example, toluene, and thelike). The reaction takes place at a temperature of about 50 to about120° C., and takes about 1 to about 24 hours to complete.

Finally, a compound of Formula 17 can be prepared by reaction of4-chloro-2-(methylthio)pyrimidine with tert-butyl cyanoacetate in thepresence of a suitable base (for example, sodium hydride and the like)and a suitable solvent (for example, DMSO and the like), at atemperature of about 25 to about 80° C. The reaction takes from about 1to about 24 hours to complete.

Compounds of Formula 11b, which are a subset of compounds of Formula 11in which M is a halogen, can be prepared by proceeding as in thefollowing Reaction Scheme VI:

in which R₇ and R₅₀ are as defined in the Summary of the Invention. Acompound of Formula 11b, in which M is a halogen, can be prepared byreacting a compound of Formula 20 with a suitable diazotization reagentsystem (for example, nitrous acid in conjunction with a copper (I)halide salt, sodium nitrite in conjunction with p-toluenesulfonic acidand potassium iodide. The reaction takes place at a temperature of about0 to about 80° C., and takes from about 1 to about 6 hours to complete.Alternatively, treatment of a compound of Formula 20 with adiazotization reagent (for example, sodium nitrite and the like) in thepresence of a carboxylic acid (for example, trifluoroacetic acid and thelike) at 0 to 40° C. for a period of about 0.5 to about 6 hours,followed by treatment with a basic aqueous solution (for example aqueouspotassium carbonate solution and the like) provides a compound ofFormula 21 (such a compound may exist in tautomeric forms). Subsequenttreatment of a compound of Formula 21 with a chlorinating agent (forexample, phosphorous oxychloride and the like), optionally in thepresence of a base such as N,N-dimethylaniline or DIPEA, and optionallyin the presence of an inert solvent such as acetonitrile or toluene, andan additive such as DMF, at a temperature of about 50 to about 110° C.,and for a time of about 1 to about 72 hours, provides a compound ofFormula 11b, in which M is chlorine.

A compound of Formula 20 can in turn be prepared by reaction of acompound of Formula 22 with guanidine or a guanidine salt (for example,guanidine hydrochloride or guanidine carbonate), optionally in thepresence of a base (for example, lithium hydroxide and the like) in asuitable solvent (for example, sec-butanol, NMP and the like), at atemperature from about 50 to about 180° C., and for a time of about 2 toabout 48 hours.

A compound of Formula 22 can be prepared by reaction of a compound ofFormula 23 with DMF DMA or Bredereck's reagent, optionally in thepresence of a co-solvent such as DMF, at a temperature of about 50 toabout 150° C. The reaction takes about 1 to about 24 hours to complete.

A compound of Formula 23 in which R₅₀ is chloro, bromo, or iodo can beprepared by reacting an amine compound of Formula 24 with a suitablediazotization reagent system (for example, nitrous acid in conjunctionwith a copper (I) halide salt, sodium nitrite in conjunction withp-toluenesulfonic acid and potassium iodide, or isoamylnitrite inconjunction with boron trifluoride-THF, iodine, potassium iodide, andacetonitrile). The reaction takes place at a temperature of about 0 toabout 80° C., and takes from about 1 to about 6 hours to complete.

A compound of Formula 24 can in turn be prepared by reacting a compoundof Formula 25 with an organometallic reagent such as methyllithium,methyllithium-lithium bromide complex, or a methylmagnesium halidereagent, in an inert solvent such as THF, ether, or cyclopropyl methylether, at a temperature of about 0 to about 100° C., followed bytreatment with an aqueous quenching solution. The reaction takes fromabout 2 to about 48 hours to complete.

A compound of Formula 25 can be prepared by reacting a compound ofFormula 26 with a compound of Formula 3, as described for ReactionScheme I. Alternatively, a compound of Formula 25 can be prepared from acompound of Formula 27, in which the two R″ groups taken together forman acid-labile protecting group (for example, an imine such asbenzylidene or a carbamate such as t-butylcarbamate). The reaction iscarried out by treatment of a compound of Formula 27 with an aqueousacidic reagent (for example, concentrated hydrochloric acid and thelike) in a solvent such as ethanol, at a temperature of about 25 toabout 100° C. Preferably the two R″ groups, taken together, form animine such as benzylidine.

A compound of Formula 27 can in turn be prepared by reacting a compoundof Formula 28 with a compound of Formula 29, in which the two R″ groupstaken together form an acid-labile protecting group (for example, animine such as benzylidene or a carbamate such as t-butylcarbamate). Thereaction is performed in a solvent (for example toluene, methanol, orethanol) at a temperature of about 25 to about 120° C., optionally inthe presence of a catalyst such as DMAP, and takes about 1 to about 24hours to complete. Optionally, the reaction is performed in an inertsolvent (for example, THF and the like) in the presence of a base (forexample n-butyllithium and the like) at a temperature of about −80 toabout 25° C., over a time of about 0.5 to about 12 hours.

A compound of Formula 29 can be prepared by methods known to thoseskilled in the art. For example, a compound of Formula 29, in which thetwo R″ groups taken together form a benzylidene group, can be preparedby reaction of benzaldehyde with a mono-substituted hydrazine R₇NHNH₂,in a solvent such as ethanol or toluene, for about 1 to about 24 hours,and at a temperature of about 25 to about 120° C. Alternatively, thereaction can be carried out using a mono-substituted hydrazine salt (forexample, a hydrochloride salt or an oxalate salt, and the like), inconjunction with a base (for example sodium acetate or triethylamine).

Compounds of Formula 8 or Formula 8a can be prepared as described in thefollowing Reaction Scheme VII:

in which R₂, R₃, R₄, R₅, and Y are as defined in the Summary of theInvention, M is a leaving group (for example, iodo, bromo, chloro,trifluoromethanesulfonyloxy, and the like), and each R′ can be, forexample hydrogen, methyl, and the like, or the two R′ groups can bejoined together to form a cyclic boronate ester. The two R₉₀ groups caneach be hydrogen, or the two R₉₀ groups, taken together, can represent asuitable nitrogen protecting group (for example, one R₉₀ can be hydrogenand the other can be BOC). Compounds of Formula 8 or Formula 8a can beprepared by reaction of a compound of Formula 50 or Formula 50a,respectively, with a diboron compound (for example,bis(pinacolato)diboron and the like) in the presence of a suitabletransition metal catalyst (for example PdCl₂(dppf)),) and a suitablebase (for example, potassium acetate and the like) in a suitable solvent(for example, toluene, dioxane and the like). The reaction proceeds in atemperature range of about 20° C. to about 120° C. and can take up toabout 24 hours to complete. A compound of Formula 50 can in turn beprepared by sulfonylation of a compound of Formula 51 as described forReaction Scheme II. It will be appreciated by one skilled in the artthat compounds of Formula 51 or Formula 50a, for example,3-bromoanilines, or N—BOC-protected 3-bromoanilines, can be prepared bya variety of methods, including, but not limited to, those describedfurther in the Examples below.

A compound of Formula 70 can be prepared as described in the followingReaction Scheme VIII:

in which R_(8b) is as defined in the Summary of the Invention and R₅₅ isselected from a C₁₋₄alkyl or halo-substituted-C₁₋₄alkyl. A compound ofFormula 70 can be prepared by treatment of a compound of Formula 71 witha suitable reducing system (for example, hydrogenation over a palladiumcatalyst, and the like) in a suitable solvent (for example, ethanol orethyl acetate) at a temperature of about 25 to about 75° C., over aperiod of about 0.5 to about 12 hours.

A compound of Formula 71 can in turn be prepared by a two step processconsisting of conversion of the hydroxy group of a compound of Formula73 to a suitable leaving group, followed by displacement with azideanion. For the first step, suitable reagents include phosphoroustribromide, or methanesulfonyl chloride in combination with a suitablebase such as triethylamine. The reactions are carried out in a suitablesolvent (for example, dichloromethane, and the like), at a temperatureof about 0 to about 50° C. For the second step, the displacement iscarried out with an azide reagent (for example, sodium azide, and thelike) in a suitable solvent (for example, DMF or DMSO) at a temperaturefrom about 25 to about 150° C. The reaction takes from about 1 to about24 hours to complete. Alternatively, the transformation can be carriedout in one step by treatment of a compound of Formula 73 with aphosphine reagent (for example, triphenylphosphine, and the like) and anazodicarboxylate reagent (for example, diethylazodicarboxylate and thelike) in the presence of hydrazoic acid (formed in situ from an azidesalt, such as sodium azide, and an acid). The reaction takes place at atemperature of about −80° C. to about 75° C., and takes about 1 to about24 hours to complete.

A compound of Formula 73 can be prepared by treatment of a compound ofFormula 74 with a chloroformate (for example, methyl chloroformate andthe like) or an alternative alkoxycarbonylation reagent such asdi-tert-butyldicarboxylate. The reaction takes place in an inert solvent(for example, dichloromethane and the like) at a temperature of about−80 to about 25° C., and takes about 1 to about 12 hours to complete. Abase such as triethylamine may optionally be used. The reaction can alsobe performed in a two-phase system consisting of a solvent such as THFor dioxane, and an aqueous basic solution, such as aqueous sodiumbicarbonate solution, at a temperature of about 25° C., for a period ofabout 2 to about 16 hours.

A compound of Formula 70 can also be prepared as described in thefollowing Reaction Scheme IX:

in which R_(8b) is as defined in the Summary of the Invention, R₅₅ isselected from a C₁₋₄alkyl or halo-substituted-C₁₋₄alkyl, and R₅₆ is asuitable protecting group, for example, benzyloxycarbonyl (CBz). Acompound of Formula 70 can be prepared by deprotection of a compound ofFormula 75. For example, a compound of Formula 70 can be prepared bytreatment of a compound of Formula 75, in which R₅₆ is Cbz, with asuitable reducing system (for example, hydrogenation over a palladiumcatalyst, and the like) in a suitable solvent (for example, ethanol,methanol, MTBE, or ethyl acetate) at a temperature of about 25 to about75° C., optionally in the presence of an acid such as hydrogen chloride,over a period of about 0.5 to about 12 hours. Alternatively, thedeprotection can be carried out under transfer hydrogenation conditions,using a suitable hydrogen donor such as formic acid, ammonium formate,or 1,4-cyclohexadiene. A compound of Formula 75 can in turn be preparedby reaction of a compound of Formula 76 with a chloroformate (forexample, methyl chloroformate and the like) or an alternativealkoxycarbonylation reagent such as di-tert-butyldicarboxylate. Thereaction takes place in an inert solvent (for example, dichloromethaneand the like) at a temperature of about −80 to about 25° C., and takesabout 1 to about 24 hours to complete. A base such as triethylamine mayoptionally be used. The reaction can also be performed in a two-phasesystem consisting of a solvent such as THF or dioxane, and an aqueousbasic solution, such as aqueous sodium bicarbonate solution, at atemperature of about 25° C., for a period of about 2 to about 16 hours.A compound of Formula 76 can be prepared by protection of a compound ofFormula 77. The protection conditions can be chosen to providepreferentially or substantially the single isomer of Formula 76.Alternatively, conditions can be chosen in which little or no preferencefor formation of the single isomer of Formula 77 is shown, but in whicha compound of Formula 76 can nevertheless be isolated in sufficientpurity for further use. Means of obtaining such purity can includecrystallization of either the free base or a salt. A compound of Formula76, in which R₅₆ is Cbz, can be prepared by treatment of a compound ofFormula 77 with benzyl chloroformate. The reaction takes place in aninert solvent (for example, dichloromethane and the like) at atemperature of about −80 to about 25° C., and takes about 1 to about 24hours to complete. A base such as triethylamine may optionally be used.The reaction can also be performed in a two-phase system consisting of asolvent such as THF or dioxane, and an aqueous basic solution, such asaqueous sodium bicarbonate solution, at a temperature of about 25° C.,for a period of about 2 to about 24 hours. A salt of a compound ofFormula 77 can be used, in conjunction with a base such as triethylamineor sodium carbonate, as input in place of the free base.

It will be appreciated by one skilled in the art that a compound ofFormula 70 can be either a single enantiomer or a mixture ofenantiomers, and that a single enantiomer compound of Formula 70 can beobtained by starting with an appropriate single enantiomer compound ofFormula 74 or Formula 77.

Detailed examples of the synthesis of a compound of Formula Ia can befound in the Examples, infra.

Additional Processes for Making Compounds of the Invention

A compound of the invention can be prepared as a pharmaceuticallyacceptable acid addition salt by reacting the free base form of thecompound with a pharmaceutically acceptable inorganic or organic acid.Alternatively, a pharmaceutically acceptable base addition salt of acompound of the invention can be prepared by reacting the free acid formof the compound with a pharmaceutically acceptable inorganic or organicbase.

Alternatively, the salt forms of the compounds of the invention can beprepared using salts of the starting materials or intermediates.

The free acid or free base forms of the compounds of the invention canbe prepared from the corresponding base addition salt or acid additionsalt from, respectively. For example a compound of the invention in anacid addition salt form can be converted to the corresponding free baseby treating with a suitable base (e.g., ammonium hydroxide solution,sodium hydroxide, and the like). A compound of the invention in a baseaddition salt form can be converted to the corresponding free acid bytreating with a suitable acid (e.g., hydrochloric acid, etc.).

Compounds of the invention in unoxidized form can be prepared fromN-oxides of compounds of the invention by treating with a reducing agent(e.g., sulfur, sulfur dioxide, triphenyl phosphine, lithium borohydride,sodium borohydride, phosphorus trichloride, tribromide, or the like) ina suitable inert organic solvent (e.g. acetonitrile, ethanol, aqueousdioxane, or the like) at 0 to 80° C.

Prodrug derivatives of the compounds of the invention can be prepared bymethods known to those of ordinary skill in the art (e.g., for furtherdetails see Saulnier et al., (1994), Bioorganic and Medicinal ChemistryLetters, Vol. 4, p. 1985). For example, appropriate prodrugs can beprepared by reacting a non-derivatized compound of the invention with asuitable carbamylating agent (e.g., 1,1-acyloxyalkylcarbanochloridate,para-nitrophenyl carbonate, or the like).

Protected derivatives of the compounds of the invention can be made bymeans known to those of ordinary skill in the art. A detaileddescription of techniques applicable to the creation of protectinggroups and their removal can be found in T. W. Greene, “ProtectingGroups in Organic Chemistry”, 3^(rd) edition, John Wiley and Sons, Inc.,1999.

Compounds of the present invention can be conveniently prepared, orformed during the process of the invention, as solvates (e.g.,hydrates). Hydrates of compounds of the present invention can beconveniently prepared by recrystallization from an aqueous/organicsolvent mixture, using organic solvents such as dioxin, tetrahydrofuranor methanol.

Compounds of the invention can be prepared as their individualstereoisomers by reacting a racemic mixture of the compound with anoptically active resolving agent to form a pair of diastereoisomericcompounds, separating the diastereomers and recovering the opticallypure enantiomers. While resolution of enantiomers can be carried outusing covalent diastereomeric derivatives of the compounds of theinvention, dissociable complexes are preferred (e.g., crystallinediastereomeric salts). Diastereomers have distinct physical properties(e.g., melting points, boiling points, solubilities, reactivity, etc.)and can be readily separated by taking advantage of thesedissimilarities. The diastereomers can be separated by chromatography,or preferably, by separation/resolution techniques based upondifferences in solubility. The optically pure enantiomer is thenrecovered, along with the resolving agent, by any practical means thatwould not result in racemization. A more detailed description of thetechniques applicable to the resolution of stereoisomers of compoundsfrom their racemic mixture can be found in Jean Jacques, Andre Collet,Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John WileyAnd Sons, Inc., 1981.

In summary, the compounds of Formula I can be made by a process, whichinvolves:

(a) that of reaction schemes I to IX; and

(b) optionally converting a compound of the invention into apharmaceutically acceptable salt;

(c) optionally converting a salt form of a compound of the invention toa non-salt form;

(d) optionally converting an unoxidized form of a compound of theinvention into a pharmaceutically acceptable N-oxide;

(e) optionally converting an N-oxide form of a compound of the inventionto its unoxidized form;

(f) optionally resolving an individual isomer, for example stereoisomer,of a compound of the invention from a mixture of isomers;

(g) optionally converting a non-derivatized compound of the inventioninto a pharmaceutically acceptable prodrug derivative; and

(h) optionally converting a prodrug derivative of a compound of theinvention to its non-derivatized form.

Insofar as the production of the starting materials is not particularlydescribed, the compounds are known or can be prepared analogously tomethods known in the art or as disclosed in the Examples hereinafter.

One of skill in the art will appreciate that the above transformationsare only representative of methods for preparation of the compounds ofthe present invention, and that other well known methods can similarlybe used.

EXAMPLES

The present invention is further exemplified, but not limited, by thefollowing intermediates and examples that illustrate the preparation ofcompounds of Formula I according to the invention.

Abbreviations used are as follows: benzyloxycarbonyl (Cbz);tert-butoxycarbonyl (BOC); Cell proliferation (CP); dichloromethane(DCM); N,N-di-isopropylethylamine (DIPEA);[1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)(PdCl₂(dppf)); 1,2-dimethoxyethane (DME); N,N-dimethyl acetamide (DMA);N,N-dimethylaminopyridine (DMAP); N,N-dimethyl formamide (DMF);N,N-dimethyl formamide dimethylacetal (DMF DMA); dimethylsulfoxide(DMSO); ethyl acetate (EtOAc); high pressure liquid chromatography(HPLC); isopropyl acetate (iPrOAc); methanesulfonyl (Ms);2-methyltetrahydrofuran (2-methylTHF); N-methylpyrolidinone (NMP);tetrahydrofuran (THF); thin layer chromatography (TLC); andpara-toluenesulfonic acid (pTsOH).

Intermediate. Cyano-(2-methylthio-pyrimidin-4-yl)-acetic acid tert-butylester

To a suspension of sodium hydride (7.15 g, 179 mmol, 60% in oil) in DMSO(100 mL) was added tert-butyl cyanoacetate (24.8 g, 170 mmol) at 23° C.After the evolution of hydrogen ceased, 4-chloro-2-methylthiopyrimidine(13.7 g, 85 mmol) was added. The reaction was heated at 80° C. for 16 h.The reaction mixture was then cooled to room temperature and quenchedwith ice-cooled saturated ammonium chloride (300 mL). The solid wasfiltered and washed with water (2×200 mL). 300 mL of hexane was added tothe solid and the suspension was heated at 60° C. for 1 h and thencooled to room temperature. The solid was filtered and washed withhexane to afford the title compound; 1H NMR 400 MHz (CDCl₃) δ 7.82 (d,1H), 6.74 (d, 1H), 2.63 (s, 3H), 2.61 (s, 1H), 1.52 (s, 9H); MS m/z:266.0 (M+1).

Intermediate. (2-Methylthio-pyrimidin-4-yl)-acetonitrile

To a solution of Intermediate cyano-(2-methylthio-pyrimidin-4-yl)-aceticacid tert-butyl ester (5.3 g, 20 mmol) in anhydrous toluene (100 mL) wasadded p-toluenesulfonic acid (800 mg). The mixture was heated to refluxfor 8 h, cooled to room temperature and extracted with ethyl acetate.The organic layer was washed with 1N aqueous sodium hydroxide solutionand brine, dried over sodium sulfate, filtered and concentrated. Thecrude product was purified by silica gel chromatography (3:1hexanes/ethyl acetate eluant) to afford the title compound: ¹H NMR 400MHz (CDCl₃) δ 8.56 (d, 1H), 7.12 (d, 1H), 3.84 (s, 2H), 2.57 (s, 3H); MSm/z: 166.0 (M+1).

Intermediate. 4-(2-(Methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine

Step 1. 3-(Dimethylamino)-2-(2-(methylthio)pyrimidin-4-yl)acrylonitrile.N,N-Dimethylformamide dimethyl acetal (30 mL) was added to Intermediate(2-Methylthio-pyrimidin-4-yl)-acetonitrile (2.62 g, 15.7 mmol) and themixture was heated at 100° C. for 16 h. The cooled reaction mixture wasconcentrated, and the residue was used without further purification.

Step 2. 4-(2-(Methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine. A mixtureof crude 3-(dimethylamino)-2-(2-(methylthio)pyrimidin-4-yl)acrylonitrilefrom the previous step (entire amount) and hydrazine monohydrate (2.36mL, 47 mmol) in anhydrous ethanol (75 ml) was heated at 80° C. for 16hours. The reaction mixture was cooled to room temperature andconcentrated. The reaction mixture was partitioned between ethyl acetateand brine. The organic layer was separated and washed with brine, driedover sodium sulfate, filtered and concentrated. The crude product waspurified by silica gel chromatography (2 to 5% methanol indichloromethane eluant) to afford4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine: ¹H NMR 400 MHz(DMSO-d₆) δ 11.9 (s, 1H), 8.3 (s, 1H), 7.87 (s, 1H), 7.23 (s, br 1H),6.43 (s, 1H), 5.74 (s, 1H), 2.53 (s, 3H); MS m/z: 208.0 (M+1).

Intermediate.I-Isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine

Intermediate 4-(2-(Methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine (10.0g, 40 mmol) was dissolved in THF (200 mL), followed by addition of2-iodopropane (6.3 mL, 63 mmol) and sodium methoxide (25% weightsolution in methanol, 14.3 ml, 63 mmol). The mixture was heated at 50°C. with stirring under a nitrogen atmosphere for 3 d, then wasconcentrated under vacuum. The residue was taken up in ethyl acetate(200 mL) and washed with aqueous potassium carbonate solution and brine,then dried over sodium sulfate, filtered, and concentrated to provide abrown residue. The residue was chromatographed on silica gel(hexane/ethyl acetate eluant) to provide1-isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine as asolid; ¹H NMR 400 MHz (CDCl₃) δ 8.23 (d, J=5.6 Hz, 1H), 7.60 (s, 1H),6.83 (d, J=5.6 Hz, 1H), 5.23 (d, J=2.8 Hz, 2H), 4.21-4.25 (m, 1H), 2.50(s, 3H), 1.37 (d, J=6.8 Hz, 6H); MS m/z: 250.1 (M+1).

Similarly prepared were:1-Ethyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine and1-Methyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine.

Intermediate.4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine

A mixture of Intermediate1-isopropyl-4-(2-(methylthio)pyrimidin-4-yl)-1H-pyrazol-3-amine (4.0 g,16.0 mmol), isopentylnitrite (13.2 g, 112 mmol), and methylene iodide(30 mL) was heated at 100° C. for 3 h. Removal of the volatiles undervacuum provided a dark residue, which was purified by silica gelchromatography (2:1 hexane/ethyl acetate eluant) to afford the titlecompound as a solid; MS m/z: 361.1 (M+1).

Similarly prepared were:4-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine;4-(3-Iodo-1-methyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine; and4-(3-iodo-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine.

Intermediate.4-(3-Iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine

A solution of 4-(3-iodo-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine (270mg, 0.85 mmol) and p-toluenesulfonic acid monohydrate (32 mg, 0.17 mmol)in 3,4-dihydro-2H-pyran (1 ml) was heated at 60° C. for 5 h. The cooledmixture was diluted with ethyl acetate, and the mixture was washed withwater and brine, and then dried over sodium sulfate, filtered, andconcentrated. The residue was chromatographed on silica gel (10% ethylacetate in hexanes eluant) to provide the title compound. MS (m/z):402.7 (M+1).

Intermediate.4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylsulfonyl)pyrimidine

To a solution of Intermediate4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylthio)pyrimidine (4.51 g,12.5 mmol) in dichloromethane (60 mL) at 0° C. was addedm-chloroperbenzoic acid (3.65 g, 77% purity, 16.3 mmol). The mixture wasstirred at 0° C. under nitrogen for 3 h, then was diluted with ethylacetate and washed with aqueous potassium carbonate solution and brine.The organic layer was dried over magnesium sulfate, filtered, andconcentrated to provide4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-2-(methylsulfonyl)pyrimidine as asolid. MS m/z: 393.0 (M+1).

Similarly prepared were:4-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)-2-(methylsulfonyl)pyrimidine; and4-(3-Iodo-1-methyl-1H-pyrazol-4-yl)-2-(methyl sulfonyl)pyrimidine.

Intermediate. 1-Benzylidene-2-isopropylhydrazine

To a round-bottom flask containing anhydrous sodium acetate (8.2 g, 0.1mol) in 125 ml 50% ethanol was added isopropylhydrazine HCl salt (11.1g, 0.1 mole) and benzaldehyde (10.6 g, 0.1 mole). The mixture wasstirred at rt for 20 h. The reaction was extracted with ether (3×250ml). The organic layers were combined and washed with aqueous sodiumbicarbonate solution and brine and dried over sodium sulfate.Filtration, concentration, and co-evaporation with toluene (3×) providedthe title compound as an oil. MS m/z 163.3 (M+1).

Similarly prepared were: 1-Benzylidene-2-ethylhydrazine starting fromethylhydrazine oxalate, using methanol and triethylamine in place ofethanol and sodium acetate, respectively; and1-Benzylidene-2-methylhydrazine starting from methylhydrazine, usingmethanol as solvent, with no added base.

Intermediate.2-((2-Benzylidene-1-isopropylhydrazinyl)methylene)-malononitrile

A solution of Intermediate 1-benzylidene-2-isopropylhydrazine (12.9 g,0.079 mol) in 200 ml anhydrous THF was cooled in a dry ice/acetone bathunder an argon atmosphere. To this solution was added n-butyllithium(1.6 M in hexanes, 66 ml, 0.106 mol) via syringe. Following completionof the addition, the mixture was stirred at dry-ice temperature for anadditional 5 minutes. A solution of (2-(ethoxymethylene)malononitrile(13.6 g, 0.11 mol) in THF (30 ml) was added. The mixture was stirred atdry-ice temperature for 0.5 h, and then was quenched with saturatedaqueous sodium bicarbonate solution. The quenched reaction mixture wasallowed to warm to rt and was extracted with ethyl acetate. The organiclayers were combined, dried over sodium sulfate, filtered, andconcentrated. The residue was suspended in ethanol with sonication. Theresulting precipitate was collected via filtration and washed with asmall amount of cold ethanol to obtain2-((2-benzylidene-1-isopropylhydrazinyl)methylene)malononitrile as ayellow solid. The mother liquor was concentrated and the residuepurified by silica gel column chromatography (1:1 hexanes/ethyl acetateeluant) to afford additional2-((2-benzylidene-1-isopropylhydrazinyl)methylene)malononitrile. MS m/z239.2 (M+1).

Intermediate.2-((2-Benzylidene-1-ethylhydrazinyl)methylene)-malononitrile

To a solution of 2-(ethoxymethylene)malononitrile (15.2 g, 0.124 mole)in 100 ml toluene was added Intermediate 1-benzylidene-2-ethylhydrazine(18.4 g, 0.124 mole). The mixture was allowed to stand at roomtemperature, and a precipitate formed. The reaction mixture was stirredfor an additional 16 h. The precipitate was collected on a filter andwashed with a small amount of cold ethanol to obtain2-((2-benzylidene-1-ethylhydrazinyl)methylene)malononitrile as a solid.

Similarly prepared was2-((2-Benzylidene-1-methylhydrazinyl)-methylene)malononitrile.

Intermediate. 3-Amino-1-isopropyl-1H-pyrazole-4-carbonitrile

A mixture of Intermediate2-((2-benzylidene-1-isopropylhydrazinyl)-methylene)malononitrile (9.42g, 40 mmol), concentrated hydrochloric acid (5 ml), and ethanol (50 ml)was heated at reflux for 20 min. The reaction mixture was concentratedand ether (50 ml) was added. The mixture was sonicated, then the upperether layer was discarded. To the residue was added 20 ml of 5N aqueoussodium hydroxide solution and the mixture was extracted withdichloromethane (3×). The organic layers were combined, dried oversodium sulfate, filtered, and concentrated. The residue was purified bysilica gel column chromatography (1:1 hexanes/ethyl acetate eluant) toafford 3-amino-1-isopropyl-1H-pyrazole-4-carbonitrile as a brown solid.¹H NMR (400 MHz, DMSO-d₆) δ 8.09 (s, 1H), 5.51 (s, 2H), 4.22 (m, 1H),1.31 (d, J=7 Hz, 6H); MS m/z 151.2 (M+1).

Similarly prepared were: 3-amino-1-ethyl-1H-pyrazole-4-carbonitrile; and3-amino-1-methyl-1H-pyrazole-4-carbonitrile.

Intermediate. 1-(3-Amino-1-isopropyl-1H-pyrazol-4-yl)ethanone

To a solution of Intermediate3-amino-1-isopropyl-1H-pyrazole-4-carbonitrile (5.29 g, 36.5 mmol) in200 ml anhydrous THF at 0° C. was added methylmagnesium bromide solution(3 M in ether, 56.5 ml, 0.17 mol). The reaction mixture was heated atreflux for 4 h. The mixture was cooled to 0° C. and was then quenchedwith 10% aqueous hydrochloric acid to neutral pH. The reaction wasextracted with a large amount of 9:1 dichloromethane/isopropanol. Theorganic layers were combined and dried over sodium sulfate, filtered,and concentrated. The crude product was purified by silica gel columnchromatography (1:1 ethyl acetate/hexanes to 9:1 ethyl acetate/methanoleluant) to afford the title compound as a light brown solid. ¹H NMR (400MHz, DMSO-d₆) δ 8.18 (s, 1H), 5.62 (s, 2H), 4.23 (m, 1H), 2.20 (s, 3H),1.37 (d, J=7 Hz, 6H); MS m/z 168.2 (M+1).

Similarly prepared were: 1-(3-Amino-1-ethyl-1H-pyrazol-4-yl)ethanone;and 1-(3-Amino-1-methyl-1H-pyrazol-4-yl)ethanone.

Intermediate. 1-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)ethanone

To a solution of Intermediate1-(3-amino-1-isopropyl-1H-pyrazol-4-yl)ethanone (3.97 g, 24 mmol) andp-TsOH.H₂O (9.07 g, 48 mmole, 2 eq) in 150 ml of acetonitrile at 0° C.was added dropwise a solution of sodium nitrite (2.97 g, 43 mmole, 1.8eq) and potassium iodide (8.0 g, 48 mmol, 2.0 eq) in 20 ml water. Themixture was stirred at this temperature for 10 minutes and then allowedto warm to rt and stirred for 3 h. The mixture was concentrated and thendiluted with water and neutralized with aqueous sodium carbonatesolution to pH 9 to 10. The mixture was extracted with ethyl acetate(3×). The combined organic layers were washed with sodium thiosulfatesolution, dried over sodium sulfate, filtered, and concentrated. Theresidue was purified by silica gel column chromatography (1:1hexanes/ethyl acetate eluant) to provide the title compound as a lightbrown solid. ¹H NMR (400 MHz, DMSO-d₆) δ 8.51 (s, 1H), 4.53 (m, 1H),2.37 (s, 3H), 1.41 (d, J=7 Hz, 6H); MS m/z 279.1 (M+1).

Similarly prepared were: 1-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)ethanone; and1-(3-Iodo-1-methyl-1H-pyrazol-4-yl)ethanone.

Intermediate.3-(Dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)prop-2-en-1-one

A mixture of Intermediate 1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)ethanone(5.0 g, 18.0 mmol) and N,N-dimethyformamide dimethyl acetal (50 mL) washeated at 155° C. for 20 h. The mixture was concentrated under vacuum toprovide the crude title compound. MS m/z 334.0 (M+1).

Similarly prepared were:3-(Dimethylamino)-1-(3-iodo-1-ethyl-1H-pyrazol-4-yl)prop-2-en-1-one; and3-(Dimethylamino)-1-(3-iodo-1-methyl-1H-pyrazol-4-yl)prop-2-en-1-one.

Intermediate. 4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-amine

A mixture of crude Intermediate3-(dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)prop-2-en-1-one(4.0 g, 12.0 mmol), guanidine hydrochloride (2.63 g, 27.6 mmol), lithiumhydroxide (635 mg, 27.6 mmol), and sec-butanol (50 ml) was heated withstirring in a sealed reaction vessel at 110° C. for 20 h. The cooledreaction mixture was concentrated, then water was added and the mixturewas extracted with ethyl acetate. The combined extracts were dried oversodium sulfate, filtered, and concentrated. Ethyl acetate was added tothe solid residue, and the mixture was sonicated. The solid product wascollected on a filter, rinsed with ethyl acetate, and dried to give thetitle compound as a tan solid. MS m/z 330.0 (M+1).

Similarly prepared were:4-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidin-2-amine; and4-(3-Iodo-1-methyl-1H-pyrazol-4-yl)pyrimidin-2-amine.

Intermediate. 4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ol

Sodium nitrite (314 mg, 4.55 mmol) was added in portions to a stirredmixture of Intermediate4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-amine (500 mg, 1.52mmol) and trifluoroacetic acid (15 ml) at 0° C. The mixture was allowedto warm to rt and was stirred for 1 h, then the solvent was removedunder vacuum. The crude mixture was diluted with ethyl acetate andwashed with saturated aqueous potassium carbonate solution and brine toprovide the title compound as a solid. MS m/z 331.0 (M+1).

Similarly prepared were:4-(3-Iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidin-2-ol; and4-(3-Iodo-1-methyl-1H-pyrazol-4-yl)pyrimidin-2-ol.

Intermediate. 2-Chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine

A solution of Intermediate4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ol (438 mg, 1.33 mmol)in phosphorous oxychloride (10 ml) was heated at 110° C. for 16 h. Themixture was concentrated under vacuum, then aqueous sodium bicarbonatesolution was carefully added and the mixture was extracted with ethylacetate. The combined organic extracts were dried over sodium sulfate,filtered, and concentrated to provide the title compound as a yellowsolid. MS m/z 349.0 (M+1).

Similarly prepared were:2-Chloro-4-(3-iodo-1-ethyl-1H-pyrazol-4-yl)pyrimidine; and2-Chloro-4-(3-iodo-1-methyl-1H-pyrazol-4-yl)pyrimidine.

Intermediate. (S)-Methyl 1-hydroxypropan-2-ylcarbamate

To a solution of (S)-alaninol (10 g, 130 mmol) and sodium bicarbonate(32.8 g, 390 mmol) in THF-H₂O (1:1, 650 mL) at 0° C. was added dropwisemethyl chloroformate (11.4 mL, 143 mmol). The mixture was stirred andallowed to warm to rt over 4 h, and was then extracted with ethylacetate. The organic layer was washed with 1N aqueous sodium hydroxidesolution and brine, and was then dried over sodium sulfate, filtered andconcentrated to give the crude title compound, which was used withoutfurther purification. MS m/z 134.1 (M+1).

Similarly prepared were: (R)-methyl 1-hydroxypropan-2-ylcarbamate, using(R)-alinol in place of (S)-alaninol; and (S)-1,1-Dimethylethyl1-hydroxypropan-2-ylcarbamate using di-t-butyl-dicarbonate in place ofmethyl chloroformate.

Intermediate. (S)-Methyl 1-azidopropan-2-ylcarbamate

To a solution of Intermediate (S)-methyl 1-hydroxypropan-2-ylcarbamate(2.65 g, 20 mmol) and triethylamine (7.0 ml, 50 mmol) in anhydrousdichloromethane (100 mL) was added methanesulfonyl chloride (1.91 mL,23.9 mmol). The mixture was stirred at rt for 3 h and was then extractedwith ethyl acetate. The organic layer was washed with 1N sodiumhydroxide solution and brine, and was then dried over sodium sulfate,filtered and concentrated. The crude mesylate product was then dissolvedin dry DMF (70 mL) and sodium azide (5.2 g, 80 mmol) was added. Themixture was heated with stirring at 80° C. for 2 h. The cooled reactionmixture was concentrated and the residue was partitioned between ethylacetate and brine. The organic layer was separated and washed withbrine, dried over sodium sulfate, filtered and concentrated. The residuewas purified by silica gel chromatography (8:1 hexane/ethyl acetateeluant) to afford (S)-methyl 1-azidopropan-2-ylcarbamate. MS m/z 159.1(M+1).

Similarly prepared were: (R)-Methyl 1-azidopropan-2-ylcarbamate; and(S)-1,1-dimethylethyl 1-azidopropan-2-ylcarbamate.

Intermediate. (S)-Methyl 1-aminopropan-2-ylcarbamate

To a solution of Intermediate (S)-methyl 1-azidopropan-2-ylcarbamate(2.86 g, 18.2 mmol) in ethyl acetate (200 ml) was added 10% palladium oncarbon (wet, 286 mg). The flask was degassed and refilled with hydrogengas (balloon, 1 atmosphere), and the mixture was stirred at rt for 16hours. The reaction mixture was filtered through a pad of celite andwashed with ethyl acetate. The filtrate was concentrated to give crude(S)-methyl 1-aminopropan-2-ylcarbamate, which was used without furtherpurification. ¹H NMR 400 MHz (CDCl₃) δ 4.79 (s, 1H), 3.71-3.65 (m, 1H),3.66 (s, 3H), 2.75 (dd, 1H), 2.65 (dd, 1H), 1.14 (d, 3H); MS m/z 133.1(M+1).

Similarly prepared were: (R)-Methyl 1-aminopropan-2-ylcarbamate; and(S)-1,1-dimethylethyl 1-aminopropan-2-ylcarbamate.

Intermediate. (S)-Methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate

A solution of Intermediate2-chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidine (1.4 g, 4.01mmol), (S)-methyl 1-aminopropan-2-ylcarbamate (0.8 g, 6 mmol) andtriethylamine (2.8 ml, 20 mmol) in isopropanol (30 ml) and dioxane (20mL) was heated in a sealed vessel at 125° C. for 48 hours. The cooledmixture was concentrated under vacuum, and aqueous sodium bicarbonatewas added to the residue. The mixture was extracted with ethyl acetate,and the combined extracts were dried over sodium sulfate, filtered, andconcentrated. The crude residue was purified by silica gel columnchromatography (1:2 hexanes/ethyl acetate eluant) to provide the titlecompound as a white solid. MS m/z 445.0 (M+1).

Similarly prepared were: (R)-Methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate;(S)-tert-butyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate;3-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propanenitrile;4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-N-methylpyrimidin-2-amine; andN¹-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-yl)-N²,N²-dimethylethane-1,2-diamine.

Intermediate. N-(3-bromo-2,4-difluorophenyl)propane-1-sulfonamide

A solution of 3-bromo-2,4-difluoroaniline (4.16 g, 20 mmol, EP184384),n-propanesulfonyl chloride (4.6 ml, 40 mmol), pyridine (8 ml), DMAP (97mg), and DCM (100 ml) was stirred at rt for 16 h. Aqueous sodiumbicarbonate solution was added and the mixture was extracted with ethylacetate. The organic layer was washed with aqueous sodium bicarbonateand brine. The crude product was purified by silica gel chromatography(8:1 to 3:1 hexanes/ethyl acetate eluant) to give the title compound. MSm/z 313.9 (M+1).

Intermediate. 3-Fluoro-4-iodopyridin-2-amine

A solution of 2-amino-3-fluoropyridine (1.0 g, 8.9 mmol) in anhydrousTHF (40 ml) was treated dropwise at −78° C. with n-butyllithium (1.6 Min hexanes, 13.9 ml, 22.3 mmol). Following the addition, the mixture wasstirred at −78° C. for 1 h, then a solution of iodine (10.2 g, 40.1mmol) in THF (20 ml) was added. The mixture was extracted with ethylacetate, and the organic extracts were washed with sodium thiosulfate,sodium bicarbonate and brine. The organic phase was dried over sodiumsulfate and concentrated. The crude product was purified by silica gelchromatography (8:1 to 2:1 hexanes/ethyl acetate eluant) to provide thetitle compound. MS m/z 238.9 (M+1).

Similarly prepared was: 3-chloro-4-iodopyridin-2-amine.

Intermediate. 3-Bromo-2,5,6-trifluoroaniline

To a microwave tube was added 1-bromo-2,3,4,5-tetrafluorobenzene (1.0 g)and 28% aqueous ammonium hydroxide (5 mL). The mixture was heated undermicrowave irradiation at 150° C. for 2 hrs, then the mixture was pouredinto water and extracted with hexane. The organic layer was separated,dried with MgSO₄, then concentrated. The residue was purified by silicagel flash chromatography (9:1 hexanes/ethyl acetate eluant) afford thetitle compound as a colorless liquid. ¹H NMR (400 MHz, CDCl₃) δ 6.75 (m,1H), 3.95 (br s, 2H) ppm; MS m/z: 226, 228 (M+H)⁺.

Intermediate. 2,4-Dibromo-3,6-dichloroaniline

A mixture of 2,5-dichloroaniline (0.2 g), N-bromosuccinimide (0.48 g)and THF (20 mL) was stirred at rt for 2 hrs. The solvent was removed,and the residue was purified by silica gel flash chromatography (8:2hexanes/ethyl acetate eluant) to afford the title compound. MS m/z: 318(M+H)⁺.

Intermediate. 1,3-Dibromo-2,5-dichlorobenzene

A stirred mixture of 2,4-dibromo-3,6-dichloroaniline (5.0 g), tert-Butylnitrite (3.3 g) and EtOH (50 mL) was heated in a sealed tube at 50° C.for 2 hrs. The mixture was concentrated and the residue was purified bysilica gel flash chromatography (hexanes eluant) to afford the titlecompound. MS m/z: 303 (M+H)⁺.

Intermediate. 3-Bromo-2-chloro-5-fluoroaniline

Step 1. 3-bromo-2-chloro-N-(diphenylmethylene)-5-fluoroaniline. Amixture of 2,6-dibromo-4-fluoro-1-chlorobenzene (865 mg, 3 mmol),benzophenone imine (0.61 ml, 3.6 mmol), Pd₂(dba)₃ (137 mg, 0.15 mmol),sodium t-butoxide (432 mg, 4.5 mmol), (S)-BINAP (280 mg, 0.45 mmol), andtoluene (30 ml) was heated at 80° C. for 16 h. The mixture was extractedwith ethyl acetate and the combined organic phase washed with brine. Theorganic phase was dried over sodium sulfate and concentrated. The crudeproduct was purified by silica gel flash chromatography (40:1 to 20:1hexanes/ethyl acetate eluant) to provide the title compound as a powder.MS m/z 388.9 (M+1).

Step 2. 3-Bromo-2-chloro-5-fluoroaniline. A solution of3-bromo-2-chloro-N-(diphenylmethylene)-5-fluoroaniline (1.16 g) in THF(20 ml) was treated with 2N hydrochloric acid (1.5 ml, 1 eq) and themixture was stirred at rt for 2 h. The mixture was extracted with ethylacetate and the combined organic phase washed with brine. The organicphase was dried over sodium sulfate and concentrated. The crude residuewas chromatographed on silica gel (40:1 to 15:1 hexanes/ethyl acetateeluant) to provide the title compound, contaminated by benzophenone. MSm/z 223.9 (M+1).

Similarly prepared were: 3-bromo-2,5-dichloroaniline;3-bromo-2-chloro-5-methylaniline; and 3-bromo-2,5-difluoroaniline.

Intermediate. 3-Bromo-5-chloro-2-fluorobenzoic acid

A solution of 2-bromo-4-chloro-1-fluorobenzene (4.31 g, 20 mmol) wasadded dropwise to a −78° C. solution of LDA in THF (prepared fromdiisopropylamine (3.38 ml, 24 mmol) and n-BuLi (1.6 M, 13.1 ml, 21mmol)). The mixture was stirred at −78° C. for 1 h, and then wastransferred slowly (˜30-60 min) via cannula to a stirred −78° C. mixtureof dry ice and THF (40 ml). The mixture was stirred at −78° C. for 1 h,and then was allowed to warm to rt (gas evolution). The mixture wasconcentrated, and was then treated with 50 ml of 1 N sodium hydroxidesolution. The mixture was extracted with ethyl acetate (discarded). Theaqueous layer was acidified with 1N hydrochloric acid, and then wasextracted with chloroform (3×400 ml). The chloroform extract was driedover sodium sulfate, filtered, and concentrated to provide the crudetitle compound. The product was contaminated with a small amount of theisomeric product 2-bromo-6-chloro-3-flurobenzoic acid. ¹H NMR for titlecompound 3-Bromo-5-chloro-2-fluorobenzoic acid: (400 MHz, CDCl₃) δ 7.93(dd, 1H, J=2.8, 5.6 Hz), 7.79 (dd, 1H, J=2.8, 5.6 Hz) ppm.

Similarly prepared were: 3-bromo-2,6-difluorobenzoic acid; and3-bromo-2-fluoro-5-methylbenzoic acid.

Intermediate. Tert-butyl 3-bromo-5-chloro-2-fluorophenylcarbamate

A solution of crude Intermediate 3-bromo-5-chloro-2-fluorobenzoic acid(2.03 g, 8 mmol), diphenylphosphoryl azide (2.07 mL, 9.6 mmol, 1.2 eq),and DIPEA (1.67 mL, 9.6 mmol, 1.2 eq) in 1:1 t-butanol/toluene (25 ml)was heated at 110° C. for 36 h. The mixture was concentrated, thenpartitioned between ethyl acetate and water. The organic layer waswashed with brine, and was then dried over sodium sulfate, filtered, andconcentrated. The crude product was purified by silica gelchromatography (30:1 to 10:1 hexanes/ethyl acetate eluant) to providethe title compound. MS m/z: 267.8 (M+H)+. (M-^(t)Bu).

Similarly prepared were: tert-butyl 3-bromo-2,6-difluorophenylcarbamate;and tert-butyl 3-bromo-2-fluoro-5-methylphenylcarbamate.

Intermediate. 3-Bromo-5-chloro-2-fluoroaniline

A solution of tert-butyl 3-bromo-5-chloro-2-fluorophenylcarbamate (900mg, 2.78 mmol) in DCM/TFA (1:1, 20 mL) was stirred at rt for 1 h. Thereaction mixture was concentrated, then the residue was taken up inethyl acetate and washed with aqueous sodium bicarbonate and brine. Theorganic layer was dried over sodium sulfate, filtered, and concentratedto provide the crude title compound (736 mg). MS m/z 223.9 (M+1).

Intermediate. 5-bromo-3-methoxy-2-methylaniline

A heterogeneous reaction mixture of 4-bromo-2-methoxy-6-nitrotoluene(500 mg, 2.032 mmol), acetic acid (20 ml), and iron (1135 mg, 20.32mmol) was stirred at rt for 24 hr. Ethyl acetate was added, then themixture was filtered through celite and the filtrate concentrated. Theresidue was partitioned between ethyl acetate and saturated aqueoussodium bicarbonate solution. The aqueous layer was extracted withfurther ethyl acetate. The combined organic phases were washed withwater and brine, then dried over sodium sulfate, filtered andconcentrated to afford the title compound. MS m/z: 218.0 (M+H)⁺.

Intermediate. Tert-butyl5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate

A mixture of Intermediate tert-butyl3-bromo-5-chloro-2-fluorophenylcarbamate (1.45 g, 4.46 mmol),bis(pinacolato)diboron (1.7 g, 6.69 mmol), potassium acetate (1.53 g,15.6 mmol), PdCl₂(dppf)-CH₂Cl₂ (163 mg, 0.22 mmol), and dioxane (100 ml)was heated in a sealed tube at 100° C. for 16 h. The crude reactionmixture was taken up in ethyl acetate and washed with aqueous sodiumbicarbonate solution and brine. The organic layer was dried over sodiumsulfate, filtered, and concentrated. The crude compound was diluted withhot hexane (600 mL), heated to 65° C. for 30 min. then cooled to roomtemperature. The brown mixture was filtered through Celite and thefilter cake was washed with hexanes. The combined filtrates wereconcentrated to afford the crude title compound as a yellow oil. MS m/z233 (M-pinacol-tBu).

Similarly prepared were:5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;tert-butyl2,6-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate;N-(2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propane-1-sulfonamide;2-(2-fluoro-3-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane;2,5-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;2-chloro-5-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;2,5-dichloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;2-chloro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;tert-butyl2-fluoro-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate;3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine;2,3,6-trifluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine;3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline;3-methoxy-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline.

Intermediate.2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

A mixture of Intermediate2-(2-fluoro-3-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (500mg), 10% palladium on carbon (50 mg) and ethyl acetate (20 ml) wasstirred under 1 atmosphere of hydrogen for 16 h. The mixture was spargedwith nitrogen and filtered. The filtrate was concentrated to provide thetitle compound, which was used without further purification. MS m/z237.1 (M+1).

Example 1 MethylN-[(2S)-1-[(4-{3-[2,6-difluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate(Compound 7 in table 1)

A mixture of crude IntermediateN-(2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propane-1-sulfonamide(854 mg), Intermediate (S)-methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(350 mg, 90% pure), tetrakis(triphenylphosphine)palladium(0) (90 mg), 2M aqueous sodium carbonate solution (6 ml), toluene (50 ml), and ethanol(6 ml) was heated at 80° C. for 16 h. The cooled mixture was extractedwith ethyl acetate and the combined organic extracts were washed withbrine. The organic phase was dried over sodium sulfate and concentrated.The crude product was purified by silica gel chromatography (70:1 to40:1 DCM/methanol eluant) to provide the title compound.

Example 2 MethylN-[(2S)-1-({4-[3-(2-chloro-5-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate(Compound 15 in table 1)

Step 1. (S)-Methyl1-(4-(3-(3-amino-2-chloro-5-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.A mixture of crude Intermediate2-chloro-5-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (214 mg), Intermediate (S)-methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(68 mg, 0.14 mmol), tetrakis(triphenylphosphine)-palladium(0) (16 mg), 2M aqueous sodium carbonate solution (3 ml), toluene (18 ml), and ethanol(3 ml) was heated at 85° C. for 16 h. The mixture was extracted withethyl acetate and the combined organic extracts were washed with brine.The organic phase was dried over sodium sulfate and concentrated. Thecrude product was purified by silica gel chromatography (60:1 to 40:1DCM/methanol eluant) to provide the title compound (46 mg) contaminatedwith (S)-methyl1-(4-(1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.MS m/z 462.1 (M+1).

Step 2. S)-Methyl1-(4-(3-(2-chloro-5-fluoro-3-(methanesulfonamido)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.A mixture of the aniline product of step 1 (46 mg), pyridine (2 ml),triethylamine (1 mL), DCM (4 ml), and methanesulfonyl chloride (23 ul,0.3 mmol) was stirred at rt for 16 h. The crude reaction mixture wasconcentrated, then the residue was taken up in a mixture of toluene (9ml), ethanol (1 ml), sodium carbonate (2 g), and water (10 ml). Thestirred mixture was heated at 85° C. for 16 h. Workup as for Step 1provided the crude product, which was purified by silica gelchromatography (60:1 to 40:1 DCM/methanol eluant) to provide the titlecompound.

Example 3 MethylN-[(2S)-1-[(4-{3-[5-chloro-2-fluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate(Compound 1 in table 1)

Step 1. (S)-Methyl1-(4-(3-(5-chloro-2-fluoro-3-(tert-butoxycarbonylamino)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.A mixture of crude Intermediate tert-butyl5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate(2.0 g), Intermediate (S)-methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(600 mg, 1.34 mmol), tetrakis(triphenylphosphine)palladium(0) (150 mg,0.13 mmol), 2 M aqueous sodium carbonate solution (6.7 ml, 13.5 mmol),toluene (80 mL), and ethanol (6 mL) was heated at 80° C. for 16 h. Themixture was extracted with ethyl acetate and the combined organicextracts were washed with brine. The organic phase was dried over sodiumsulfate and concentrated. The crude product was purified by silica gelchromatography (80:1 to 60:1 DCM/methanol eluant) to provide the titlecompound contaminated with triphenylphosphine oxide. MS m/z 562.1 (M+1).

Step 2. (S)-Methyl1-(4-(3-(3-amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.A solution of partially-purified (S)-methyl1-(4-(3-(5-chloro-2-fluoro-3-(tert-butoxycarbonylamino)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate (1.15 g) in DCM (50 mL) and TFA (20 mL) wasstirred at rt for 1 h. The solvents were removed, then aqueous sodiumbicarbonate solution was added. The mixture was extracted with ethylacetate, and then the combined organic extracts were washed with sodiumbicarbonate and brine. The organic phase was dried over sodium sulfateand concentrated. The product was obtained by silica gel chromatography(60:1 to 40:1 DCM/methanol eluant); MS m/z 462.1 (M+1).

Step 3. MethylN-[(2S)-1-[(4-{3-[5-chloro-2-fluoro-3-(propane-1-sulfonamido)phenyl]-1-(propan-2-yl)-1H-pyrazol-4-yl}pyrimidin-2-yl)amino]propan-2-yl]carbamate.A mixture of (S)-methyl1-(4-(3-(3-amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(41 mg, 0.09 mmol), triethylamine (1 ml), and DCM (4 ml) was treatedwith propanesulfonyl chloride (40 mg, 0.27 mmol). The mixture wasstirred at rt for 16 h. The crude mixture was concentrated, and theresidue was taken up in a mixture of toluene (9 ml), ethanol (1 ml),sodium carbonate (2 g), and water (10 ml). The stirred mixture washeated at 85° C. for 16 h. Workup as for Step 1 provided the crudeproduct, which was purified by silica gel chromatography (60:1 to 40:1DCM/methanol eluant) to provide the title compound.

Example 4 MethylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(oxan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate(Compound 33 in table 1) and MethylN-[(2S)-1-({14-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate(Compound 31 in table 1)

Step 1.5-Chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)aniline.Prepared according to the procedure of Example 3, step 1, starting fromintermediate4-(3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)-2-(methylthio)pyrimidineand intermediate5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline.MS m/z 420.0 (M+1).

Step 2.N-(5-Chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)phenyl)methanesulfonamide)andN-(5-chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)phenyl)-N-(methylsulfonyl)methanesulfonamide.To a solution of5-chloro-2-fluoro-3-(4-(2-(methylthio)pyrimidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)aniline(233 mg, 0.55 mmol) and triethylamine (2 mL) in dichloromethane (10 mL)was added methanesulfonyl chloride (0.13 mL, 1.66 mmol). The mixture wasstirred at room temperature for 16 h, providing a mixture of the titlecompounds (LCMS analysis). Ethyl acetate was added and the mixture waswashed with water and brine, then the organic layer was dried oversodium sulfate, filtered, and concentrated to provide the crude titlemixture, which was used without further purification. MS m/zmono-sulfonamide 498.0 (M+1); bis-sulfonamide 576.0 (M+1)

Step 3. MethylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(oxan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate.The crude product mixture from Step 2 was dissolved in THF-H₂O (1:1, 30mL) and treated at rt with Oxone® (1.68 g, 2.75 mmol). The mixture wasstirred at rt for 16 h, then ethyl acetate was added. The organic layerwas washed with aqueous sodium bicarbonate solution, water, and brine,then was dried over sodium sulfate, filtered, and concentrated. Thecrude residue was dissolved in NMP (5 mL) and treated with intermediate(S)-methyl 1-aminopropan-2-ylcarbamate (146 mg, 1.1 mmol) and sodiumcarbonate (233 mg, 2.2 mmol). The mixture was heated with stirring at110° C. for 16 h. The cooled reaction mixture was diluted with ethylacetate, and was then washed with water and brine. The organic layer wasdried over sodium sulfate, filtered, and concentrated. The residue waschromatographed on silica gel (2% methanol in dichloromethane eluant) toprovide the title compound.

Step 4. MethylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate.To a solution of methyl(2S)-1-(4-(3-(5-chloro-2-fluoro-3-(methylsulfonamido)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(94 mg, 0.16 mmol) in MeOH (15 mL) was added concentrated hydrochloricacid (0.5 mL). The mixture was stirred at rt for 16 h. Aqueous sodiumbicarbonate was added to adjust the pH to 9 and the mixture wasextracted with ethyl acetate. The organic layer was washed with aqueoussodium bicarbonate and brine. The crude product was purified by silicagel chromatography (30:1 to 15:1 dichloromethane/methanol eluant) toprovide the title compound.

Example 5 MethylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate(Compound 9 in table 1)

Methanesulfonyl chloride (0.277 mL, 3.57 mmol) was added to a solutionof (S)-methyl1-(4-(3-(3-amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(550 mg, 1.2 mmol) in DCM (30 mL) and pyridine (10 mL), and the mixturewas stirred at rt for 16 h. Aqueous sodium bicarbonate solution wasadded, and the mixture was extracted with ethyl acetate and washed withbrine. The organic phase was dried over sodium sulfate and concentrated.The crude product was purified by silica gel chromatography (60:1 to40:1 DCM/methanol) to provide the title compound. An alternativesynthesis is described in example 6, infra.

Also isolated from the reaction mixture were: MethylN-[(2S)-2-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propyl]carbamate(Compound 32 in table 1);N-{3-[4-(2-aminopyrimidin-4-yl)-1-(propan-2-yl)-1H-pyrazol-3-yl]-5-chloro-2-fluorophenyl}methanesulfonamide(Compound 30 in table 1).

Example 6 MethylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate(Compound 9 in table 1)

Step 1. 1-Benzylidene-2-isopropylhydrazine. Into a reactor equipped witha mechanical stirrer, a thermometer, and an addition funnel undernitrogen purge were charged isopropylhydrazine hydrogenchloride salt(712 g, 6.43 mol), sodium acetate (528 g, 6.43 mol), and 50% ethanol(4500 mL). The mixture was stirred at 20° C. for 5 min. Benzaldehyde(683 g, 6.43 mol) was added while maintaining the batch temperaturebelow 23° C. The mixture was stirred at 20° C. over 20 h. Toluene (6500mL) was added and stirring was maintained for 5 min. The organic layerwas separated. Saturated aqueous sodium bicarbonate solution (4800 mL)was slowly added to the vigorously stirred organic layer (Note: the pHof the aqueous layer was ˜8.0). The organic layer was separated andwashed with saturated aqueous sodium bicarbonate solution (3000 mL).Then the organic layer was separated and concentrated under vacuum(50→20 torr) at 40° C. to give the title compound as a yellow oil (usedwithout further purification).

Step 2.2-((2-Benzylidene-1-isopropylhydrazinyl)methylene)-malononitrile. To aflask equipped with a mechanical stirrer, a thermometer, and an additionfunnel under nitrogen purge were charged (ethoxyethylidine)malononitrile(755 g, 6.18 mol), DMAP (150 g, 1.23 mol), and ethanol (6400 mL). Themixture was stirred to give a dark orange solution and an endotherm from20° C. to 12° C. was observed. 1-Benzylidene-2-isopropylhydrazine (1101g, crude) was added slowly over 15 min to give an exotherm to 32° C. andan orange suspension. The orange suspension was heated to 50° C. andheld at 50° C. for 30 min to give a dark brown suspension. Ethanol (3200mL) was added to the mixture and the mixture was cooled to 20° C. andheld at 20° C. for 1 h. The slurry was filtered and the solid cake wasrinsed with ethanol (3000 mL). The solid was collected and dried undervacuum at 40° C./5 torr for 3 h to furnish the title compound as ayellow solid. HPLC purity >99%.

Step 3. 3-Amino-1-isopropyl-1H-pyrazole-4-carbonitrile. To a flaskequipped with a mechanical stirrer, a thermometer, a condenser, and anaddition funnel under nitrogen purge were charged2-((2-benzylidene-1-isopropylhydrazinyl)methylene)-malononitrile (632.6g, 2.65 mol), MeOH (2.5 L), and cone. (12 N) HCl (329.0 mL, 3.94 mol).The mixture was heated to 63° C. and held at 63° C. for 30 min to givean orange solution. The mixture was cooled to 15° C. Heptane (4 L) andMTBE (1 L) were added and the mixture stirred for 5 min. Then water (7.5L) was added in a stream over 30 min at 15° C. to 25° C. The mixture wasstirred 10 min at 25° C. after the water addition was complete. Theheptane/MTBE layer was separated. The aqueous layer was washed with a(4:1 v/v) heptane/MTBE mixture (2×5 L) by stirring each wash for 10 minat 25° C. The layers were separated. Solid sodium chloride (1 Kg) wasadded to the aqueous layer. Saturated aqueous potassium carbonatesolution was then added to the aqueous layer slowly to control the CO₂evolution and to adjust the pH to ˜9.0. The aqueous layer was thenextracted twice with CH₂Cl₂ (1×2.2 L, 1×800 mL). The combined CH₂Cl₂layers were dried over MgSO₄, filtered and the filtrate was concentratedunder vacuum (200 torr) at a bath temperature of 30° C. until theresidual weight was ˜1 Kg. Heptane (6.0 L) was slowly added (over ˜20min) to the CH₂Cl₂ solution with stirring and a slurry was formed. Themixture was concentrated under vacuum (60 torr) at an internaltemperature of 25° C. until the residual volume was ˜6.2 L. The slurrywas cooled to 15° C. and held at this temperature for 10 min. Theproduct was filtered and the solid was washed with heptane (1 L). Thesolid was dried under vacuum (5 torr) at 30° C. for 4 h to furnish thetitle compound as a yellow solid. HPLC purity >99%.

Step 4. 1-(3-Amino-1-isopropyl-1H-pyrazol-4-yl)-ethanone. To a flaskfitted with a mechanical stirrer, thermometer, reflux condenser,heating/cooling capacity, addition funnel, and nitrogen inlet/outlet wascharged 3-amino-1-isopropyl-1H-pyrazole-4-carbonitrile (274 g, 1.82mole) and cyclopentyl methyl ether (2600 mL) at 20° C. under nitrogen.The suspension was cooled to −10° C. 1.5 M Methyllithium/lithium bromidecomplex in diethyl ether solution (6.0 L, 9.00 mol) was added dropwiseover 2.5 h at −10° C. to 0° C. When the methyllithium addition wascomplete the reaction suspension was quickly warmed to 5° C. to 10° C.and held in this temperature range for 1 h. The mixture was cooled to 0°C. and 2N HCl (6.0 L) was added dropwise at 5-10° C. The (upper) organiclayer was separated and extracted with 2N HCl (500 mL). The combinedaqueous layers were stirred at rt over 16 h. The mixture was cooled to15° C. and basified with 50% NaOH (260.0 g) to give a pH of ˜11.0. Themixture was extracted with CH₂Cl₂ (1×2.0 L, 1×1 L). The combined CH₂Cl₂layers were dried over MgSO₄, filtered and concentrated under vacuum togive a yellow solid (278 g). The solid was dissolved in EtOAc (750 mL)upon heating to 65° C. The solution was cooled to rt and a slurry wasobtained. Heptane (1500 mL) was added slowly over 40 min at rt. Theslurry was cooled to −10° C. and held at −10° C. for 30 min. The slurrywas filtered and the filter cake was rinsed with heptane (300 mL). Thesolid was dried under vacuum (5 torr) at 40° C. for 3 h to give thetitle compound as a light brown solid. HPLC purity >99%. M.P. 136-139°C.

Step 5. 1-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-ethanone. A flaskequipped with a mechanical stirrer, a thermometer, and an additionfunnel under nitrogen purge was charged with1-(3-amino-1-isopropyl-1H-pyrazol-4-yl)-ethanone (250.0 g, 1.49 mol) andacetonitrile (3725 mL). The mixture was cooled to −20° C. BF₃.THF (313.1g, 2.23 mol) was added dropwise while keeping the internal temp. <−10°C. Isoamyl nitrite (227.5 g, 1.94 mol) was added dropwise while keepinginternal temp. <−10° C. The mixture was allowed to warm to 10° C. andwas stirred at 10° C. for 30 min. The mixture was added in a thin streamto a flask containing a mixture of I₂ (28.5 g, 0.112 mol), KI (371.9 g,2.24 mol) and acetonitrile (1160 mL) at 10-15° C. with vigorousstirring. The addition caused the evolution of nitrogen gas and a slightexotherm. The mixture was stirred at rt for 30 min. HPLC showed nodiazonium intermediate left. Then sodium bisulfite (157.1 g, 1.51 mol)in 8% sodium chloride solution (4360 mL) was added at 15-20° C. Themixture was basified with saturated potassium carbonate to pH ˜8.5. Thetop acetonitrile layer was separated and concentrated under vacuum togive an oily residue. The oil was dissolved in iPrOAc (2770 mL) andwashed with saturated aqueous sodium carbonate solution (1100 mL). TheiPrOAc layer was separated and concentrated to a residual volume of ˜1.5L. A suspension was obtained. To the suspension was added heptane (5.5L) over 30 min at 20° C. The suspension was stirred for 10 min at 20° C.after the heptane addition was complete. The slurry was filtered and thesolid was rinsed with heptane (1 L) and then dried at 20° C. undervacuum (5 torr) for 16 h to give the title compound. MP: 90-92° C.

Step 6.3-(Dimethylamino)-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-prop-2-en-1-one.To a flask equipped with a mechanical stirrer, a thermometer, and anaddition funnel under nitrogen purge were charged1-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-ethanone (640 g, 2.30 mol) andDMF (6.4 L). The resulting orange solution was heated to 120° C.Bredereck's reagent (598.6 g, 3.43 mol) was added in one portion. Theaddition caused the batch temperature to decrease to 114° C. and thesolution turned darker orange in color. The mixture was stirred at 120°C. for 20 min. The mixture was cooled to rt and then concentrated at 5mmHg at 60° C. to give an oily residue. The residue was dissolved iniPrOAc (2400 mL) by warming to 74° C. The mixture was cooled to 35° C.and stirred to obtain a slurry. Heptane (6000 mL) was added at 35° C. tort over 1 h. The mixture was cooled to −15° C. and filtered and thesolid was dried under vacuum at 40° C. for 3 h to give the titlecompound as a solid. HPLC purity >98%. MP: 106-109° C.

Step 7. 4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidin-2-amine. To aflask equipped with a mechanical stirrer, a thermometer, a Dean-Starktrap, and a condenser under nitrogen purge were charged(E)-3-dimethylamino-1-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-prop-2-en-1-one(735 g, 2.2 mol), guanidine carbonate (596 g, 3.3 mol) and NMP (5200mL). The mixture was heated to 130° C. and held at 130° C. for 5 h.(Note: any low-boiling fractions were collected by the Dean-Stark trap).The mixture was cooled to 80° C. and 15% aq. sodium chloride (7500 mL)was added at 80° C. to 35° C. over ˜1 h. The product began toprecipitate approximately half way thru the aqueous sodium chlorideaddition. The mixture was further cooled to rt and held for 30 min. Thesolid product was collected by filtration and dried under vacuum at 65°C. for 16 h to give the title compound as a solid. HPLC purity >99%. MP:167-169° C.

Step 8. 4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidin-2-ol. A flaskequipped with a mechanical stirrer and thermometer under nitrogen purgewas charged with TFA (748.8 mL).4-(3-Iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidin-2-amine (300 g, 0.91mol) was added in portions as a solid while maintaining the internaltemperature below 30° C. using a cold water bath. The mixture wasstirred at rt for 10 min to obtain a solution. The mixture was cooled to20° C. and sodium nitrite (79.7 g, 1.27 mol) was added in portions over5 h at 22-28° C. with rapid stirring. (Note: some gas evolution wasobserved and there was a mild exotherm that was easily controlled usinga cool water bath). DCM (12 L) was added and the mixture was warmed to27° C. Water (4400 mL) was added (Note: gas evolution at beginning).Saturated potassium carbonate solution (˜1500 mL) was added slowly tothe mixture to basify to pH ˜9.0 (Note: A large amount of gas wasevolved). To the mixture was added a solution of sodium bisulfite (32 g,0.30 mol) in water (100 mL). The mixture was stirred at 27° C. for 15min and the pH was readjusted to ˜9.0. The DCM layer was separated andconcentrated under vacuum (200-100 mmHg) at a bath temp of 40° C. untilthe residual weight was ˜2300 g (˜1750 mL). To the residue was addedMTBE (1500 mL) at 20° C. The mixture was stirred 10 min at 20° C. andwas then filtered. The solid was dried 16 h at 30° C. under vacuum (5torr) to give the title compound as an off-white solid. HPLCpurity >99%. MP: 216-218° C.

Step 9. 2-Chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidine. To aflask fitted with a stirrer, thermometer, condenser, addition funnel andnitrogen inlet/outlet was charged4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidin-2-ol (311 g, 942 mmol).To the solid was added acetonitrile (2500 mL) at 20° C. The mixture wasstirred to give a suspension. To the suspension was added DIPEA (246.2mL, 1.41 mol), followed by DMF (218.8 mL, 2.83 mol). The resultingsuspension was stirred 5 min at 20° C. To the suspension was added POCl₃(217 g, 1.41 mol) at 20-40° C. to give an orange solution. The mixturewas warmed to 80° C. and held at 80° C. for 3 h. The mixture was cooledto 10° C. and a solution of ammonium hydroxide (622 mL of 28%) indeionized water (5550 mL) was added slowly over 1.5 h, keeping thetemperature below 20° C. After completion of the ammonium hydroxideaddition, the resulting suspension was stirred for 40 min at 10-20° C.The solid product was collected by filtration and dried overnight at 40°C. under vacuum (5 torr) to give the title compound as a brown solidHPLC purity >99%.

Step 10. (S)-Benzyl 2-(methoxycarbonylamino)propylcarbamate. To asuspension of (S)-1,2-diaminopropane dihydrochloride (50 g, 340 mmol) indichloromethane (500 mL) was added potassium carbonate (1190 mmol). Thesuspension was stirred and filtered to collect the filtrate. Thefiltrate was cooled to 0-5° C. and stirred while benzyl chloroformate(51 ml, 357 mmol) was added dropwise. Following completion of theaddition, the reaction mixture was stirred for 3 h at 0-5° C., and wasthen allowed to warm to rt and was stirred at rt overnight. To thismixture was added dropwise triethylamine (71 ml, 510 mmol) and themixture was cooled to 0-5° C. Methyl chloroformate (28 ml, 357 mmol) wasadded slowly at 0-5° C. and the mixture was allowed to warm to rt andwas stirred overnight. The mixture was poured into water. The organicvolatiles were removed under vacuum. The resulting aqueous slurry wasfiltered to collect the solids, and the filter cake was then washed withethyl acetate to provide a white solid (65 g, 92-94% HPLC purity).Multiple recrystalizations from ethyl acetate provided the titlecompound as a white solid, HPLC purity 99.5%.

Step 11. (S)-Methyl 1-aminopropan-2-ylcarbamate Hydrochloric Acid Salt.A solution of (S)-Benzyl 2-(methoxycarbonylamino)propylcarbamate inmethanol was hydrogenated over a 5% palladium/C catalyst at 50-60 psi.The reaction mixture was filtered and the filtrate was concentratedunder vacuum to give a colorless oil. 60 g of the colorless oil wasdissolved in 200 mL of anhydrous dichloromethane and the solution wascooled to 0-5° C. in an ice-water bath. A solution of HCl in methanol(ca. 75 mL) was added dropwise until the pH of the solution was <1. Theresulting suspension was stirred at 0-5° C. for 30 min, then the solidwas collected via filtration. The solid was washed with dichloromethaneand then with hexanes to give the title compound as a white solid.

Step 12. 3-Bromo-5-chloro-2-fluorobenzaldehyde. A solution of2,2,6,6-tetramethylpiperidine (327 g, 98%, 2.274 mol) and THF (1.9 L,HPLC grade) was cooled to −75° C. (dry ice-methanol bath) under an argonatmosphere. 1.6 M n-BuLi/hexane solution (1.47 L, 2.35 mol) was addedslowly into the mixture at −72 to −67° C. over 1 h. The mixture wasstirred at −72 to −67° C. for 30 min to give a pale yellow suspension.2-Bromo-4-chloro-1-fluorobenzene (435 g, 97%, 2.02 mol) was added slowlyinto the mixture at −72 to −67° C. over 30 min, and then the mixture wasstirred at −72 to −67° C. for an additional 30 min. Dimethylformamide(230 g, 99.5%, 3.14 mol) was added slowly into the mixture at −70 to−65° C. over 30 min and then the mixture was stirred at −70 to −65° C.for 30 min to afford a light brown solution. The cooling bath wasremoved and then saturated ammonium chloride solution (720 mL) was addedinto the batch at −60 to −30° C. over 15 min to obtain a hazy mixture. 6N hydrochloric acid was quickly added into the mixture at −30 to 10° C.over 15 min to pH 1 and then ethyl acetate (2.0 L) was added at 10 to20° C. The layers were separated and the aqueous layer was extractedwith ethyl acetate (1×300 mL). The combined organic extracts were washedwith water (1×800 mL) and brine (1×500 mL), dried over magnesiumsulfate, and filtered. The filtrate was concentrated under vacuum(60-65° C.) to give the title compound as a tan viscous oil, whichsolidified upon standing after several hours. ¹H NMR (CDCl₃): δ7.76-8.30 (m, 2H), 10.0-10.8 (br s, 1H); MS m/z 238.0 (M+1).

Step 13. 3-Bromo-5-chloro-2-fluorobenzoic acid. A stirred mixture of3-bromo-5-chloro-2-fluorobenzaldehyde (415 g), tert-butanol (1.2 L) andwater (1.2 L) was warmed to 30° C. and then potassium permanganate (335g, 2.12 mol) was added (5 portions) into the batch at 40-45° C. over 1h. The dark purple contents were heated in a step-wise fashion at 45-50°C. for 30 min, 50-55° C. for 30 min and 55-60° C. for 30 min to afford apurple-brown suspension. The reaction mixture was allowed to cool to 20°C., then saturated sodium sulfite solution was added at 22-27° C., untila negative peroxide test was obtained. Warm water (2.5 L, ˜50° C.) andsaturated sodium carbonate solution (100 mL) were added sequentiallyinto the mixture over 15 min. The dark suspension was filtered through a1 cm celite bed, and the filter cake was washed with warm water (4×1 L,˜50 C). The combined filtrate was acidified with 6 N hydrochloric acidsolution to pH 1 to obtain a yellow oily suspension. Ethyl acetate (3 L)was added into the mixture and the mixture was stirred for 10 min. The(upper) organic layer was washed with water (1.2 L), dried overmagnesium sulfate, filtered, and concentrated under vacuum (60-65° C.)to give a thick yellow suspension. Hexane (700 mL) was added into theresidue and the suspension was cooled to 5-10° C. The solid wascollected by filtration, and the filter cake was dried under vacuum (65°C.) overnight to give the title compound as a yellow solid. mp 150-152°C.; HPLC purity (225 nm): 97.5%; ¹H NMR (d₆-DMSO): δ 7.82 (s, 1H), 8.10(s, 1H), 13.82 (br s, 1H); MS m/z 254 (M+H).

Step 14. Tert-butyl 3-bromo-5-chloro-2-fluorophenylcarbamate. A mixtureof 3-bromo-5-chloro-2-fluorobenzoic acid (243 g, 97.5%, 0.935 mol),triethylamine (105 g, 99.5%, 1.02 mol), and tert-butanol (1.4 L) washeated to 74° C. A solution of diphenylphosphoryl azide (260 g, 97%,0.916 mol) in toluene (960 mL) was added slowly into the mixture at75-79° C. over 1 h (gentle refluxing). The mixture was heated slowly to83° C. over 30 min and maintained at 83-84° C. (gentle refluxing) for 1h. The contents were concentrated under vacuum (65-70° C.) to a viscousoil. Toluene (2 L) and water (1.5 L) were added sequentially into thebatch and the mixture was then stirred at 35° C. for 15 min. The aqueouslayer was discarded The organic layer was washed with saturated sodiumbicarbonate solution (400 mL) and water (400 mL). The organic layer wasconcentrated under vacuum (60-65° C.) to ˜350 mL residue (˜94% purity).10% ethyl acetate/hexane (˜1.2 L, v/v) was added into the batch and themixture was then heated at 50° C. for 15 min to give a light yellowhomogenous solution. Ethyl acetate/hexane solution was transferred ontoa premade silica gel (1.8 kg, 70-200 mesh)/hexane bed on a 4-L PyrexBüchner funnel (with coarse fritted discs, 40-60 μm, 16 cm diameter, 18cm height). t-Butyl carbamate product was eluted (by gravity) slowlywith 3-5% ethyl acetate/hexane (total volume ˜5 L, v/v) to collect thetitle compound as an off-white solid. mp 87-88° C.; HPLC purity (225nm): 97-98%; ¹H NMR (d₆-DMSO): δ 1.48 (s, 9H), 7.48-7.49 (m, 1H),7.80-7.82 (m, 1H), 9.42 (s, 1H); MS m/z 325 (M+H).

Step 15. (S)-Methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.To a 4-neck flask equipped with a mechanical stirrer, a thermometer, anda condenser under nitrogen purge were charged2-Chloro-4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)-pyrimidine (300.0 g),(S)-methyl 1-aminopropan-2-ylcarbamate hydrochloric acid salt (174.3 g),sodium carbonate (365.7 g) and DMSO (2400 mL). The mixture was heatedwith stirring for 18 h at an internal temperature of 90° C. The mixturewas cooled to 40° C. Toluene (3870 mL) was added at 37-43° C. withstirring. Water (7200 mL) was added at 37-43° C. The Toluene layer wasseparated at 37-43° C. To the Toluene layer was added 15% aqueous sodiumchloride 1 solution (3870 mL) and the pH of the aq. layer was adjustedto a pH ˜5.0 by the addition of 10% aqueous citric acid solution at37-43° C. The pH adjustment required ˜20 mL of 10% aqueous citric acid.The toluene layer was then washed with saturated aqueous sodiumbicarbonate solution (2880 mL) at 37-43° C. The toluene layer containingthe title compound was used as input in step 17.

Step 16. Tert-butyl5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate.To a flask equipped with a mechanical stirrer, thermometer, condenser,and heating mantle under nitrogen purge were charged tert-butyl3-bromo-5-chloro-2-fluorophenylcarbamate (33.0 g),bis(pinacolato)diboron (447.0 g), potassium acetate (405.6 g) andtoluene (2700 mL). The mixture was stirred at rt for 15 min andPdCl₂(dppf) (50.4 g) was added. The mixture was then heated to 108±2° C.(Note: The mixture turned dark at 50-60° C.) A solution of tert-butyl3-bromo-5-chloro-2-fluorophenylcarbamate (414 g) in toluene (1770 mL)was added at 108±2° C. over 70 min. The mixture was held at 108±2° C.for 15 h. The mixture was cooled to rt under nitrogen flow and then wasfiltered through celite. The filtrate containing the title compound wasused as input in step 17.

Step 17. (S)-Methyl1-(4-(3-(5-chloro-2-fluoro-3-(tert-butoxycarbonylamino)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate. To a flask was charged (S)-methyl1-(4-(3-iodo-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(3870 ml toluene solution, ˜382 g, 0.861 mol) and tert-butyl5-chloro-2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate(4470 ml toluene solution, ˜467.0 g, 1.26 mol). To the resulting brownsolution was added a solution of sodium carbonate (349.8 g, 3.30 mol) inwater (1400 mL). To the mixture was added PdCl₂(dppf) (34.5 g, 0.047mol). The mixture was warmed to 80° C. with stirring and was held atthis temperature for 2 h. The mixture was cooled to 40° C. and filteredthru celite. The layers in the filtrate were separated. The toluenelayer containing the title compound was used directly as input in step18.

Step 18. (S)-Methyl1-(4-(3-(3-amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.To a flask was charged (S)-methyl1-(4-(3-(5-chloro-2-fluoro-3-(tert-butoxycarbonylamino)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(˜7.3 L toluene solution, ˜483.3 g, 0.86 mol) at 20° C. under a nitrogenatmosphere. To the solution was added 12N HCl (574.3 mL, 6.95 mol) over˜20 min while maintaining the temperature below 25° C. The HCl additioncaused an exotherm from 19° C. to 24° C. The mixture was stirred at20-23° C. for 1 h. To the reaction mixture was added water (3100 mL).The mixture was stirred 10 min at 20° C. The aqueous layer was separatedand washed with 2-methylTHF (3100 mL). To the aqueous layer was slowlyadded saturated aqueous potassium carbonate solution (˜778 mL). The pHof the aqueous layer was ˜8.5. The aqueous layer was extracted with2-methylTHF (3825 mL). The 2-methylTHF layer was concentrated undervacuum (60 mmHg, 40° C.). The residue was diluted with 2-methylTHF(˜3800 mL) to provide a solution of the title compound, which was useddirectly as input in step 19. HPLC purity: 95%.

Step 19. (S)-Methyl1-(4-(3-(5-chloro-2-fluoro-3-(N-(methylsulfonyl)methylsulfonamido)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate.To a flask was charged (S)-methyl1-(4-(3-(3-amino-5-chloro-2-fluorophenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(˜3.8 L methylTHF solution, ˜396.5 g, 0.86 mol) at 20° C. To thesolution was added triethylamine (435.0 g, 4.3 mol). The solution wascooled to 0 to −5° C. To the solution was added methanesulfonyl chloride(246.0 g, 2.15 mol) dropwise with stirring over 20 min at 0 to −5° C.The mixture was allowed to warm to 18-20° C. and was held at thistemperature for 20 min. To the reaction mixture was added water (2115mL) over 30 min at 18-20° C. The mixture was stirred 10 min at 20° C.after the addition was complete. Then the pH was adjusted to between 6.0and 6.5 with 2N HCl (˜1230 mL). The pH was then adjusted to 7-7.5 usingsaturated aqueous sodium bicarbonate solution. The mixture was stirred10 min @20° C. The layers were separated. The 2-methylTHF layercontaining the title compound was used directly in step 20.

Step 20. MethylN-[(2S)-1-({4-[3-(5-chloro-2-fluoro-3-methanesulfonamidophenyl)-1-(propan-2-yl)-1H-pyrazol-4-yl]pyrimidin-2-yl}amino)propan-2-yl]carbamate.To a flask was charged (S)-methyl1-(4-(3-(5-chloro-2-fluoro-3-(N-(methylsulfonyl)methylsulfonamido)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-ylamino)propan-2-ylcarbamate(˜3.8 L methylTHF solution, ˜531.5 g, 0.86 mol). To the solution wasadded 3N aqueous sodium hydroxide (1782.8 mL, 5.34 mol) at 15-20° C.with stirring. The mixture was vigorously stirred at 20-23° C. over 30min and the stirring was stopped. The aqueous layer was discarded. 2NHCl (˜410 mL) was added to the organic layer to adjust the pH to6.0-6.5, then saturated aqueous sodium bicarbonate solution (˜300 mL)was added to adjust the pH ˜8.5. The aqueous layer was discarded. Theorganic layer was washed with 15% aqueous sodium chloride solution (2000mL). The organic layer was concentrated under vacuum (80 torr) at a bathtemperature of 45° C. to give a brown solution (780 g). The solution wasdiluted with 2-methylTHF (3500 mL), and then a suspension of PICA HP120N activated carbon (90 g, CDH858) in 2-methylTHF (1 L) was added. Theresulting black suspension was warmed to 60° C. and held at 60° C. for16 h. After the 16 h hold the Pd content was 309 ppm. The mixture wascooled to 20° C. and filtered through a pad of celite (prewetted with2-methylTHF). The reaction flask was rinsed with 2-methylTHF (500 mL).This rinse was then poured thru the filter cake of PICA/celite. To thefiltrate was added PL-TMT resin (90 g). The resulting suspension washeated with stirring to 60° C. and held at this temperature for 4 h.After 4 h at 60° C. the Pd content was 2.3 ppm. The mixture was cooledto 20° C. and stirred overnight at 20° C. The mixture was filteredthrough a pad of celite (prewetted with 2-methylTHF). The filtrate wasconcentrated under vacuum (100-80 torr) at 40-45° C. to give an orangeoil. This residual oil was dissolved in 3 L of 200 proof ethanol bywarming to 78° C. The resulting clear orange solution was then cooled to20° C. over 3 h, and a precipitate formed. The mixture was then cooledto 0° C. and held 1 h at 0° C. The mixture was filtered and the filtercake was washed with ethanol (300 mL). The solid was dried 14 h at 40°C. to give the title compound; MP: 186-189° C.

The compounds in the following table 1 were prepared similarly to theabove examples, using the appropriate starting materials:

TABLE 1 BRAF V600E bio- A375 CP chemical Physical Data Cpd IC₅₀ IC₅₀ ¹HNMR 400 MHz # Structure (μM) (μM) and/or MS (m/z) 1

0.002 ¹H NMR 400 MHz (CD₃OD) δ 8.41 (s, 1H), 8.06 (d, 1H), 7.57 (dd,1H), 7.34 (dd, 1H), 6.64 (d, 1H), 4.63 (hept, 1H), 3.62-3.68 (m, 1H),3.57 (s, 3H), 3.40-3.44 (m, 1H), 3.32-3.36 (m, 1H), 3.05 (q, 2H), 1.88(q, 2H), 1.58 (d, 6H), 1.02 (d, 3H), 0.97 (t, 3H); MS m/z 568.2 (M + 1).2

0.018 0.0008 MS m/z 522.1 (M + 1) 3

0.002 MS m/z 534.1 (M + 1) 4

0.033 0.0023 MS m/z 506.1 (M + 1) 5

0.001 0.0002 MS m/z 550.1 (M + 1) 6

0.046 0.0039 MS m/z 524.2 (M + 1) 7

0.008 0.0007 MS m/z 552.2 (M + 1) 8

0.027 MS m/z 588.2 (M + 1) 9

0.002 0.0003 ¹H NMR 400 MHz (CD₃OD) δ 8.41 (s, 1H), 8.08 (d, J = 5.6 Hz,1H), 7.57 (dd, J = 6.4, 2.6 Hz, 1H), 7.34 (dd, J = 5.6, 2.6 Hz, 1H),6.65 (brs, 1H), 4.63 (hept, J = 6.8 Hz, 1H), 3.62-3.68 (m, 1H), 3.57 (s,3H), 3.40- 3.44 (m; 1H), 3.32- 3.36 (m, 1H), 3.00 (s, 3H), 1.58 (d, J =6.8 Hz, 6H), 1.02 (d, J = 4.8 Hz, 3H).MS m/z 540.1 (M + 1) 10

0.003 0.0006 ¹H NMR 400 MHz (CD₃OD) δ 8.41 (s, 1H), 8.08 (d, 1H), 7.37(ddd, 1H), 7.08 (ddd, 1H), 6.62 (brs, 1H), 4.63 (hept, 1H), 3.63 (s,3H), 3.54-3.58 (m, 1H), 3.34-3.40 (m, 2H), 3.00 (s, 3H), 1.58 (d, 6H),1.12 (d, 3H); MS m/z 524.1 (M + 1). 11

0.003 0.0005 ¹H NMR 400 MHz (CD₃OD) δ 8.34 (s, 1H), 8.09 (d, 1H), 7.57(dd, 1H), 7.35 (dd, 1H), 6.65 (brs, 1H), 4.28 (q, 2H), 3.68-3.71 (m,1H), 3.57 (s, 3H), 3.32-3.36 (m, 2H), 3.00 (s, 3H), 1.54 (t, 3H), 1.02(d, 3H); MS m/z 526.0 (M + 1). 12

0.004 0.00095 1H (400 MHz, CDCl₃) δ 8.12 (s, 2H), 7.40 (d, 1H), 7.20 (d,1H), 7.10 (m, 1H), 6.40 (s, 1H), 5.35 (s, 1H), 5.25 (d, 1H), 4.60 (m,1H), 3.90(s, 1H), 3.65 (s, 3H), 3.30 (s, 1H), 3.00 (s, 3H), 2.40 (s,3H), 1.60 (d, 6H), 1.10 (d, 3H). MS (ESI) m/z: 521 (M + H)⁺. 13

0.006 0.0010 1H (400 MHz, CDCl₃) δ 10.10 (s, 1H), 8.60 (s, 1H), 7.75 (s,1H), 7.60 (d, 1H), 7.15 (d, 1H), 7.05 (s, 1H), 6.30 (s, 1H), 4.95 (s,1H), 4.65 (m, 1H), 3.95 (s, 1H), 3.65 (s, 3H), 3.45 (s, 1H), 3.30 (s,1H), 3.05 (s, 3H), 2.45 (s, 3H), 1.65 (d, 6H), 1.20 (d, 3H). MS (ESI)m/z: 537 (M + H)⁺. 14

0.079 MS m/z 523.1 (M + 1)¹ 15

0.004 0.0011 MS m/z 540.1 (M + 1) 16

1.4 0.024 MS m/z 507.2 (M + 1) 17

0.002 MS m/z 478.1 (M + 1) 18

0.012 ¹H NMR 400 MHz (CD₃OD) δ 8.45 (s, 1H), 8.04 (d, 1H), 7.41 (dd,1H), 6.57 (d, 1H), 4.61 (hept, 1H), 3.70- 3.74 (m, 1H). 3.58 (s, 3H),3.34-3.40 (m, 2H), 3.23 (q, 2H), 1.58 (d, 6H), 1.43 (t, 3H), 1.02 (d,3H); MS m/z 573.1 (M + 1). 19

0.093 0.0061 MS m/z 524.1 (M + 1) 20

0.007 0.0008 MS m/z 540.2 (M + 1) 21

0.045 ¹H (400 MHz, CDCl₃) δ 7.95 (s, 2H), 7.30 (s, 1H), 6.40 (s, 1H),5.15 (s, 1H), 4.50 (m, 1H), 3.70 (s, 1H), 3.52 (s, 3H), 3.20 (s, 1H),3.05 (s, 3H), 1.50 (d, 6H), 1.05 (d, 3H). MS (ESI) m/z: 542 (M + H)⁺. 22

0.006 ¹H (400 MHz, CDCl₃) δ 8.50 (s, 1H), 7.85 (s, 1H), 7.80 (s, 1H),7.30 (s, 1H), 6.40 (s, 1H), 5.0 (s, 1H), 4.62 (m, 1H), 3.90 (s, 1H),3.65 (s, 3H), 3.30 (s, 1H), 3.10 (s, 3H), 1.70 (d, 6H), 1.20 (d, 3H). MS(ESI) m/z: 557 (M + H)⁺. 23

1.15 MS m/z 488.2 (M + 1) 24

0.023 MS m/z 550.1 (M + 1) 25

0.002 MS m/z 566.1 (M + 1) 26

0.014 0.0017 MS m/z 538.2 (M + 1) 27

>20 0.39 MS m/z 532.2 (M + 1) 28

0.55 0.09 MS m/z 496.1 (M + 1) 29

0.076 0.008 MS m/z 439.0 (M + 1) 30

0.069 0.0043 MS m/z 425.1 (M + 1) 31

0.18 0.0008 MS m/z 497.9 (M + 1) 32

0.031 0.0029 MS m/z 540.1 (M + 1) 33

0.007 MS m/z 582.1 (M + 1) 34

0.013 0.0027 ¹H NMR 400 MHz (CD₃OD) δ 8.41 (s, 1H), 8.06 (d, 1H), 7.47(ddd, 1H), 7.17 (ddd, 1H), 6.59 (d, 1H), 4.62 (hept, 1H), 3.71-3.74 (m,1H), 3.58 (s, 3H), 3.34-3.40 (m, 2H), 3.05 (q, 2H), 1.88 (q, 2H), 1.58(d, 6H), 1.02 (d, 3H), 0.97 (t, 3H); MS m/z 552.1 (M + 1).

Example 122 B-Raf V600E/Mek Amplified Luminescence Proximity HomogeneousAssay (B-Raf V600E Biochemical)

B-Raf (V600E; 4 pM) and biotinylated Mek (kinase dead; 10 nM) werecombined at 2× final concentrations in assay buffer (50 mM Tris, pH 7.5,15 mM MgCl₂, 0.01% BSA and 1 mM DTT) and dispensed 10 μl per well inassay plates (Greiner white 384 well assay plates #781207) containing0.5 μl of 40× of a compound of the invention diluted in 100% DMSO. Theplate was incubated for 60 minutes at room temperature.

The B-Raf kinase activity reaction was started by the addition of 10 μlper well of 2×ATP (10 μM) diluted in assay buffer. After 3 hours, thereactions were stopped with the addition of 10 μl of stop reagent (60 mMEDTA, 0.01% Tween20). Phosphorylated product was measured using a rabbitanti-p-MEK (Cell Signaling, #9121) antibody and the Alpha Screen IgG(ProteinA) detection Kit (PerkinElmer #6760617R), by the addition of 30μl to the well of a mixture of the antibody (1:2000 dilution) anddetection beads (1:1000 dilution of both beads) in bead buffer (50 mMTris, pH 7.5, 0.01% Tween20). The additions were carried out under darkconditions to protect the detection beads from light. A lid was placedon top of the plate and the plate was incubated for 1 hour at roomtemperature. The luminescence was read on a PerkinElmer Envisioninstrument. The concentration of each compound for 50% inhibition (IC₅₀)was calculated by non-linear regression using XL Fit data analysissoftware.

Compounds of the invention, in free form or in pharmaceuticallyacceptable salt form, exhibit valuable pharmacological properties, forexample, as indicated by the in vitro tests described in thisapplication. For example, compounds of the invention preferably show anIC₅₀ in the range of 1×10⁻¹⁰ to 1×10⁻⁵ M, preferably less than 500 nM,250 nM, 100 nM and 50 nM for V600E B-Raf.

For example, IC₅₀ data for some compounds of the invention in theLuminescence Proximity Homogeneous Assay are shown in the Table, supra.

Example 123 A375 Cellular Proliferation Assay (A375 CP)

A375 is a melanoma cell line that harbors the B-Raf V600E mutation.A375-luc cells engineered to express luciferase are plated to 384-wellwhite clear bottom plates as 1,500 cells/50 μl/well in DMEM containing10% FBS. Compounds of the invention dissolved in 100% DMSO atappropriate concentrations are transferred to the cells by a robotic PinTool (100 nl). The cells are incubated for 2 days at 25° C., then 25 μlof BrightGlo™ is added to each well and the plates are read byluminescence. The concentration of each compound for 50% inhibition(IC₅₀) was calculated by non-linear regression using XL Fit dataanalysis software.

Compounds of the invention, in free form or in pharmaceuticallyacceptable salt form, exhibit valuable pharmacological properties, forexample, as indicated by the in vitro tests described in thisapplication. For example, compounds of the invention preferably show anIC₅₀ in the range of less than 500 nM, 250 nM, 100 nM and 50 nM for wildtype and V600E B-Raf.

For example, IC₅₀ data for some compounds of the invention in the A375Cellular Proliferation Assay are shown in the Table, infra.

Example 124 Immunoassays

Cells were seeded in 1640 RPMI+10% FBS at a density of 30×10³ cells per96-well in tissue culture treated plates, then incubated at 37° C. and5% CO₂ for 24 hours before compounds were added. Test compounds wereserially diluted in DMSO then added to the cells (final DMSOconcentration of 0.1%) and incubated at 37° C. and 5% CO₂ for 3 hours.pMEK and pERK levels were measured using a sandwich immunoassay kit(Meso Scale Discovery). Culture supernatants were removed and cells werelysed by the addition of cold lysis buffer (provided in kit) for 30minutes with gentle shaking. For detection of pMEK1/2 (Ser217/221) andpERK1/2 (Thr/Tyr202/204, Thr/Tyr185/187), lysates were added to theblocked antibody coated plates provided with the kits and incubatedovernight at 4° C. with shaking. Plates were washed and thephosphoproteins detected using the provided labelled antibodies and readon a Sector 6000 instrument.

Example 125 SW620 Cell Viability Assay

SW620 cells were seeded in 1640 RPMI+10% FBS at a density of 1500 cellsper 96-well in black walled, clear bottom tissue culture treated plates.Test compounds were serially diluted in DMSO then added to the cells(final DMSO concentration of 0.1%) and incubated at 37° C. and 5% CO₂for 4 days. To measure cell viability, cell plates were brought to roomtemperature, culture media was removed, and 200 μl of Cell Titer-Gloreagent (Promega, kit components mixed as per the manufacturer'sprotocol then diluted 1:2 with growth media) was added to each well.Plates were shaken for 5 minutes, then incubated at room temperature for5 minutes, and the luminescence was measured (Trilux, Perkin Elmer).

Example 126 Rat1 Soft Agarose Assay

Rat1 cells were suspended in 1% agarose (Lonza) at a density of 1000cells per 96-well. The agarose/cell mixture was allowed to solidify.Test compounds were serially diluted in DMSO then added on top of theagarose cell mixture (final DMSO concentration of 0.2%) and incubated at37° C. and 5% CO₂. After 17 days, colony growth was determined byincubating cells with alamarBlue (TREK Diagnostics) and measuring themetabolic activity with a Spectramax plate reader (Molecular Devices,Inc; absorbance measured at 562 nm).

Example 127 Liver Microsomal Clearance Assay

The in vitro microsomal clearance assay is designed to assess potentialrisks associated with hepatic metabolic stability of compounds. The testcompound (1 μM) was incubated with liver microsomes (0.5 mg/mL) fromdifferent species (mouse, rat, monkey, dog and human) and NADPH (1 mM)in 100 mM potassium phosphate buffer at 37° C. At specific reaction timepoints (0, 5, 10 and 30 minutes), reaction aliquots were removed andreactions were terminated by addition of ice cold acetonitrilecontaining mass spectrometry internal standard. The samples werecentrifuged and the supernatants were analyzed by LC-MS/MS. In vitrometabolic half-life (t_(1/2), min) and intrinsic clearance (CLint,μL/min/mg) are based on the rate and extent of metabolism of the testcompound as determined by the disappearance of the parent compound fromthe reaction mixture. These values may be scaled to predict hepaticmetabolic clearance rate (CLh, mL/min/kg) and extraction ratio (ER,expressed as a ratio of the predicted hepatic metabolic clearance tohepatic blood flow in that species). In general, compounds with highpredictedCLint or ER in vitro are considered to be at high risk forexposure-limiting metabolism in vivo.

Measured extraction ratios for some compounds of the invention are givenin the table below.

ER ER Cpd Structure Human Mouse 9

<0.21 0.48 7

0.65 0.91 4

<0.31 0.40 6

0.69 3

0.69 0.85 1

0.66 0.79

Example 128 A549 p38α MAP Kinase Bright-Glo Reporter Gene Assay

A549 cells were stably-transfected with the IL-8 promoter drivenreporter, pGL3-IL8-Luc. The cells were plated at 4×10⁵/ml into 384-wellsolid white plates (40 ul/well, 5% CD-FBS, 1×P/S, DMEM) and wereincubated overnight (18-20 hours) at 37° C. Test compounds were seriallydiluted in DMSO, then 50 nl of test solution was added to the incubation(final DMSO concentration of 0.1%). After incubating with test compoundfor 30 min, cells were stimulated with Ing/ml IL-1 beta (10 ul of 5ng/ml solution per well). Bright-Glo (25 ul/well) was added to measureluciferase expression after 7-8 hours of stimulation. IC₅₀ data for somecompounds of the invention are given in the table below.

A549 p38 RGA A375 IC₅₀ CP IC₅₀ (p38 RGA)/ Cpd Structure (μM) (μM) (A375)ratio 9

2.2 0.002 1100 32

0.69 0.031 91 29

17 0.076 220

Example 129 In Vivo Pharmacokinetics Assay

Full pharmacokinetics study: Male Balb/c mice (n=3, body weights 22-25g) or male Wistar rats (n=3, body weights 250-300 g) were administeredthe test compound intravenously via the lateral tail vein or orally viagavage. The formulation was typically a 2.5 mg/mL solution of thecompound in 75% PEG300 and 25% D5W. Six blood samples of 50 μL each werecollected serially for 24 h after dosing. The blood samples werecentrifuged to separate the plasma. Plasma samples were analyzed andquantified by LC-MS/MS.

Rapid pharmacokinetics study: Male Balb/c mice (n=3, body weights 22-25g) or male Wistar rats (n=3, body weights 250-300 g) were administeredthe test compound intravenously (IV) via the lateral tail vein or orally(PO) via gavage. The formulation was typically a 2.5 mg/mL solution ofthe compound in 75% PEG300 and 25% D5W. Six blood samples of 50 μL eachwere collected serially for 24 h after dosing. Blood samples werecentrifuged and plasma separated and pooled across the three animals ateach time point per dose route. Plasma samples were analyzed andquantified by LC-MS/MS.

For both full and rapid pharmacokinetics studies, the followingparameters were calculated by non-compartmental regression analysisusing Winnonlin 5.0 software (Pharsight, Mountain View, Calif., USA):plasma clearance (Cl), plasma maximum concentration (Cmax), plasmaarea-under-the-concentration-time-curve (AUC_(0-inf)), and percent oralbioavailability (F %).

Pharmacokinetics parameters in mouse for some compounds of the inventionare given in the table below.

Dose (mg/kg), Cl(mL/ PO AUC PO Cmax Compound # Structure PO/IV min/kg)(hrs*μM) (μM) F (%) 9

10/2 4.3 ± 0.4 30 ± 4 27 ± 4 43 ± 6 7

10/2 99 ± 13 0.24 ± 0.21 0.15 ± 0.07 8 ± 7 4

10/2 41 ± 9 3.0 ± 0.5 2.0 ± 0.5 36 ± 6 6

10/2 61 ± 6 1.1 ± 0.2 0.60 ± 0.02 15 ± 3 3

5/2 51 (n = 1) 0.20 (n = 1) 0.07 (n = 1) 6 (n = 1) 1

5/2 7.0 (n = 1) 9.7 (n = 1) 7.4 (n = 1) 46 (n = 1)

Compounds of Formula I, in free form or in pharmaceutically acceptablesalt form, exhibit valuable pharmacological properties, for example, asindicated by the in vitro and in vivo tests described in thisapplication. For example, compounds of Formula I preferably show an IC₅₀in the A375 CP cell proliferation assay in the range of 250 nM orbetter, preferably less than 200 nM, 150 nM, 100 nM and 50 nM.

The 2-(methoxycarbonylamino)-1-propyl group at R₁ provides for apreferred level of activity and selectivity over other kinases includingp38. For example, a greater than 30 fold increase in activity existsbetween compounds 9 and 29 where the A375 IC₅₀ is 2 nM and 76 nM,respectively.

The phenyl substitution pattern of compounds of Formula Ib is optimalfor metabolic stability (ER in mouse and human) with fluoro or chloro atthe R₅ position and fluoro, chloro, or methyl at the R₃ position.Compare, for example, the ER (human) in compounds 9 and 6 of <0.21 and0.69, respectively.

The combination of the 2-(methoxycarbonylamino)-1-propyl group at R₁,the R₃/R₅ substitution pattern and the methyl group at R₄ has asurprising effect on the total drug exposure (AUC). See, for example,compound 9 (compared to compounds 1, 3, 4, 6 and 7) where the AUC, at anoral dose of 10 mg/kg, is 30±4 micromolar*hrs.

Example 130 In Vivo Efficacy—14 Day A375 Mouse Xenograft Model

A375 cells were grown in sterile conditions in a 37° C. incubator with5% CO₂ for two to four weeks. The cells were cultured in RPMI-1640 mediasupplemented with 10% FBS. Cells were passaged twice weekly with 0.05%Trypsin/EDTA. On the day of implant, cells were harvested in HBSS(Hank's Balanced Salt Solution). Female Nu/Nu mice (Charles River, 10-11weeks at start of study) were implanted with 5×10⁶ A375 cells/mouse in50% matrigel, 0.2 mL SQ right flank on day 1. At 19 days post-implant,mice were randomized into 6 groups (9 mice per group), with an averagetumor volume of 215 mm³ and average body weight of 24 g. Test compoundwas dosed BID for 14 days commencing on day 19 using a dosing volume 0.2mL per dose, with compound formulated at the appropriate concentrationto attain the desired dose level. Clinical observations were made daily.Tumor volume and body weight were measured twice weekly. Endpoints: Anyindividual animal or group that exhibited body weight loss exceeding 25%of initial and/or tumor volume in excess of 3000 mm³.

Compound 9 (formulated in 20% PEG300/3% ETPGS/77% water) was dosedaccording to the above protocol using the following dosing regimen:

Group 1: Vehicle, qd×14

Group 2: 1 mg/kg Cpd 9, bid×14

Group 3: 3 mg/kg Cpd 9, bid×14

Group 4: 10 mg/kg Cpd 9, bid×14

Group 5: 20 mg/kg Cpd 9, bid×14

Tumor volume results were assessed at 14 days post-first dose. Partialresponse is defined as having tumor growth 20-50% of control tumorgrowth. Stable disease is defined as having final tumor volume within+/−20% of initial tumor size. Partial regression is defined as havingfinal tumor volume <80% of initial tumor volume.

Group 2: partial response

Group 3: stable disease

Group 4: partial regression

Group 5: partial regression

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference for allpurposes.

We claim:
 1. A compound of Formula Ia:

in which: Y is selected from N and CR₆; R₂ R₃, R₅ and R₆ areindependently selected from hydrogen, halo, cyano, C₁₋₄alkyl,halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy and halo-substituted-C₁₋₄alkoxy;with the proviso that when R₅ is fluoro and R₁ is selected fromhydrogen, —X₁R_(8a), —X₁OX₂R_(8a), —X₁C(O)NR_(8a)R_(8b),—X₁NR_(8a)X₂R_(8b), —X₁NR_(8a)C(O)X₂OR_(8b) and —X₁NR_(8a)S(O)₀₋₂R_(8b),R₃ and R₆ are not both hydrogen; R₄ is selected from —R₉ and —NR₁₀R₁₁;wherein R₉ is selected from C₁₋₆alkyl, C₃₋₈cycloalkyl,C₃₋₈heterocycloalkyl, aryl and heteroaryl; wherein said alkyl,cycloalkyl, heterocycloalkyl, aryl or heteroaryl of R₉ is optionallysubstituted with 1 to 3 radicals independently selected from halo,cyano, C₁₋₄alkyl, halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy andhalo-substituted-C₁₋₄alkoxy; and R₁₀ and R₁₁ are independently selectedfrom hydrogen and R₉; R₇ is selected from hydrogen, C₁₋₄alkyl,C₃₋₅cycloalkyl and C₃₋₅heterocycloalkyl; wherein said alkyl, cycloalkylor heterocycloalkyl of R₇ is optionally substituted with 1 to 3 radicalsindependently selected from halo, cyano, hydroxyl, C₁₋₄alkyl,halo-substituted-C₁₋₄alkyl, C₁₋₄alkoxy and halo-substituted-C₁₋₄alkoxy;or the tautomers, prodrugs, stereoisomers or pharmaceutically acceptablesalts thereof.